 Okay, so again, good morning, good afternoon, good evening, whatever time you're watching this asynchronous class of ours for clinical chemistry one. So again, I'm greeting everybody a good day. So we'll be continuing our discussion about analytical techniques, instrumentation, and also eventually on the latter part of our discussions we'll be talking about your automation. So again, I hope everybody is doing well. So if you want to follow along, we're still on chapter five of your bishop. So if you want to follow, well, I am discussing you may use your bishop chapter five. So for this morning, we'll actually begin talking about your electrophoresis. So I hope you already have finished watching spectrophotometry, AAS fluorometry, and even your chemiluminescence, okay, your luminescence for today. We're going to discuss electrophoresis in this particular video. So let's get started and proceed with our discussion. So as you all know, electrophoresis is one of the most common instrumentation right now, not only in clinical chemistry, but as we move towards molecular biology, you'll actually be more familiar and you'll be introduced to electrophoresis even more. So what do we mean by electrophoresis? So it's actually a play of words, a combination of your electro, which means your electrical charge or your electricity. Phoresis means separation through the use of your pores, where we will be talking about that later on when we talk about your stationary medium for this particular instrumentation. So to get started, what is electrophoresis? So electrophoresis is the separation of charge compound based on their electrical charge. So take, for example, you have a particular solution with different molecules in it. And remember that these molecules, they do have their own charges. So whether they are positively charged or negatively charged ions, whether they are cat ions or anions, they will be migrating through our medium and they will be separated according to their charges. They will be separated according to their charges. So electrophoresis in some of your references is also defined as the process of separating your charged constituents. So just like what I was mentioning, on your solution, we have different charged analytes or constituents where we will be separating them through the use of electrical current. We'll be separating them through the use of electrical current. So when we're talking about electrophoresis, the two most common methods of electrophoresis is your ionophoresis and your zone electrophoresis. Your ionophoresis and your zone electrophoresis. So moving forward, let's talk more about your electrophoresis. So an important thing that you need to know first, just like what I was mentioning, your analytes, they are ions with charge. So if a particular molecule or a particular substance has the ability to be both positively or negatively charged depending on the isoelectric point or the pH of a particular solution, we refer to them as amphoteric. So amphoteric, these are substances that can have either a negative charge, a zero charge or a positive charge depending on the condition. And when we're talking about your condition, you can under an encircle condition and write this beside it. When we're talking about conditions, we're talking about your isoelectric point and also your pH. We're talking about your isoelectric point and your pH. So those conditions would determine or would dictate if a amphoteric molecule or an amphoteric ion would be a positively negatively charged or a neutral molecule. So a perfect example of your amphoteric or an amphoteric molecule are your proteins. So depending on the isoelectric point and pH of a particular solution, they can be positively or negatively charged depending again on your condition. So let me just be clear when it comes to amphoteric. Your amphoteric can be positive or negative. So one at a time lang. So a particular molecule can be positive. A particular molecule can be negative according to or depending to a particular condition. That is amphoteric. Why do I have to clear that? Because we also have your zwiter ion or zwiterion. So when we talk about zwiterion or zwiterion, this is a particular molecule that has both your positive and negative charge at the same time. So that's their difference when it comes to amphoteric and zwiterion or zwiterion depending on how you would want to pronounce it. So remember that amphoteric and your zwiterion are your zwiterion. On the other hand, we also have your anions. So when we talk about anions, these are negatively charged. These are negatively charged molecule. These are negatively charged molecule that migrates to the anode. Later on, we'll discuss what are anode and cathode for you to be able to understand them. So the reason why we call them anion, because these molecules migrate to the anode. They migrate to the anode. So when we talk about anion, these are ions with negative charge. On the other hand, we also have your cat ion. Cat ions, these are positively charged ions. On the other hand, which migrate to the cathode? Again, when we say cat ion, they have a positive charge. These are positively charged ion that would migrate to the cathode. So let me just be clear between cathode and anode. Because most of the time, some of the students have mistaken that anode is negatively charged and cathode is positively charged. Actually, it's not. Your anode, these are actually your positively charged electrode. These are your positively charged electrodes. So just like in magnetism, opposite charges or opposite poles attracts. Opposite poles attract. Similarly, when it comes to your charges, opposite charges would also attract. That is the reason why your negatively charged ion would migrate or would be, yeah, they would migrate to your positively charged electrode, which is now your anode. Okay? So I hope we're clear with that. So if it is an anion, we're talking about the ion, so it is negatively charged. But if we're talking about anode and cathode, anode is positively charged electrode and your cathode is the negatively charged electrode. That is also the same reason why your cat ions or positively charged ions will be attracted or will migrate towards your cathode. So this is our very important foundational, foundational concepts when it comes to electrophoresis so that when we move along to your discussion with electrophoresis, you'd be able to understand anion, cat ions, anodes and electrodes better. Okay? Now, with that being said, okay, with that being said, I also just want to remind everybody that when it comes to electrophoresis, there are different conditions or different factors that could actually affect the mobility of your particle. So please do remember that when it comes to electrophoresis, perhaps just an advanced discussion to you guys to remember that electrophoresis and chromatography are somehow similar. Again, I'm telling, I'm saying they are similar. Similar in the sense that we are separating substances or molecules from a particular solution. So one is passive, the other one is the other one which is your electrophoresis would apply now, your electricity. Okay? So even though we have your electricity, other factors can still play into role. When I say they can still play into role, meaning to say they can still affect. Okay? They can still affect your electrophoresis. So one of those things that could actually affect the mobility of your particle are number one, the net charge of your particle. Okay? When we talk about the net charge of your particle, whether they are positively or negatively charged if they are an ion or cat ion, that would affect their mobility. So always remember that most of the time, okay, most of the time the electrode nearest to your sample well, okay, the electrode nearest to your sample well are your cathode and then the farthest ones are your anode, okay? The farthest one are your anode. So always remember that your net negative, your net charge, okay, your net charge would actually affect, okay, your net charge would affect the mobility of your particle. Aside from that, obviously, okay, we're also going to talk about your size and the shape of the particles. Remember that this molecule will pass through a porous membrane which we call your support medium, okay, or your support media. Later on, we'll discuss about that, the different components of your electrophoresis. So their size and their shape would also affect their mobility, okay? So just like, take for example, yeah, you are trying to, you have your filter, okay? You have your filter and you're trying to decant a particular solution. So the rate of decantation would also affect depending on the size of the particle, okay, the size of the particle that you would want to be removed and also the size of your, the size or your medium in itself, which is bullet number four, chemical property or physical properties of your medium, okay? So the chemical and physical properties of your medium could also affect the mobility of your particle. The strength of electric or electrical field could also affect your substances from migrating. So if you apply too low, okay, too little electrical charge or electricity on the system, then it will be slower. If you up it up a notch, if you accidentally have it to the maximum, it will also be fast, okay? It would also be the migration of your particles would also be too fast, okay? So you would want to have an electrical charger at the strength of your electrical field, a little on the middle that would allow your molecules or your particle to migrate into your medium properly. And of course, we also have your electrophoretic temperature. Always remember that when temperature is lower than the optimal temperature your particle might move slower. So just like if it's colder temperature if the colder temperature particles, the mobility of your particle will be slower. And if it is too high, okay? Or the temperature is hotter, okay? Or warmer, the particle would migrate faster than the usual. So again, we would want to have, we would want to be on the middle, the optimal temperature, optimal electrical charge to have an optimal procedure, an optimal electrophoretogram by the end of your procedure. Now having said that, okay? These are the following components of your electrophoresis. So we have your power supply, obviously we have your buffer. You also have your support medium, your sample, okay? The sample in itself or the sample well. And also the detecting system. So we'll be talking about them one by one. Although it will be brief. I just want to create a new video about electrophoresis because my previous video or my previous lecture about electrophoresis is a bit chap, a bit long. And also we're having issues when it comes to its audio, okay? So having said that, let's proceed with the first component of your electrophoresis which are your power supply. So your power supply obviously supplies a constant current or voltage in the system. So just want to highlight constant current because any fluctuation or your current might affect your electrophoretic pattern. So always remember, you can either use a UPS or an EVR, so that the voltage of your machine or the voltage of your electrophoresis would be regulated. Aside from that, your power supply since it would be providing the electrical current for the system, it would also be the one that would drive your molecules through your support medium. Again, just like what I am mentioning, your electrophoresis is a type of a kinetic diff it uses your energy already. So that it could drive the particles faster compared to a usual diffusion, okay? So if we're going to depend on diffusion that would take longer, turn around time, longer procedure compared to when you're using your electrical current already. So this is also important. Your power source is also important when we're talking about other instrumentation and other techniques not only in Glen Ken but also in other sections of the medical laboratory science such as your immunology and serology and in some cases even your blood banking and hematology, okay? So aside from that, your power supply is also known as your driving force, obviously because it would be the one that would help, okay? It would be the one that would help your that would help your ions or your particles move through your support medium. So again, remember this that the voltage depend on the ionic strength of your buffer which is another component of your electrophoresis. So why do we need to why do we need to maintain a specific voltage and why is it dependent on the ionic strength of the buffer? Remember that when it comes to your electrophoresis not only do we maintain a constant voltage or a constant electrical current but we also want to maintain a constant a constant pH, okay? A constant pH within our system. So having a having having to compute that and knowing your ionic the ionic strength of your buffer would also let you know how much or what voltage will you be using for your electrophoresis? Okay, so that is your power supply. So obviously next to your power supply of course we have your buffer. So similarly to the buffer that is in our body your buffer in electrophoresis also carry the same function. So the your buffer okay, remember that your buffer is used to provide ions that would carry the current and that would maintain the pH at a relatively constant value. So remember that buffers are substances or compounds that would maintain or that would prevent changes in your pH. Okay? So please do remember that buffer okay your buffer okay also your buffer are also ion that will enable the movement of your current and the migration of your particle. They are like conductors within your electrophoresis system. Remember that for the electricity to flow or for the electricity to migrate for the current to pass through the medium it would need your buffer. I add it would need your buffer. So remember your buffer being your being the one that would maintain your pH and at the same time it will also enable the movement. Okay? So remember your buffer the movement of your current okay? Remember had the movement of your current. So from the power supply or the power source it can move along or it can move through your support medium through the help of your buffer. Okay? Through the help of your buffer. So those are the first two components of your electrophoresis we're finished with your power supply next is some of the we're going to go through some of the important things about buffer before we move along to the next component. So remember that your pH and your ionics the ionic strength of your buffer might affect your analyte. That is why maintaining your buffer at a particular pH is very important. Okay? So remember ionic strength was mentioned a while back in the computation or in knowing how much voltage or how much electricity or how much voltage will you be applying for your system and at the same time it also gives you an idea about the pH that is again important and in your substances. Let me just let me just reiterate the importance of buffer and pH in electrophoresis. So a while back we were talking about ampoteric, correct? We're talking about ions that can either be positive or negative depending on the pH of a particular solution. So if that if there are changes or drastic changes in your pH the mobility of your proteins could also change. So unlike the one that is already expected for their movement, okay? All of all your biomolecules has already we already know their electrophoretic mobility. We usually compare it into a control. So the problem is he the problem now is that if you have problem with your pH or you have problem with your ionic strength that could affect their electrical electrophoretic mobility. So instead of being the first one to migrate they can become second or last because of problems when it comes to your pH and also your ionic strength. So that's why buffer is needed inside your electrophoretic system. So in general your buffer is also a mixture of proton donating and also proton accepting substances. So remember a simple buffer. So some are electron proton donating and proton accepting so that they can maintain pH at a constant level. So the usual buffer that we're using are actually your barbital or your verona with a pH with a pH of 8.6 or in some cases we can also use your trisborek EDTA your trisborek EDTA with a pH of 8.7. Again your trisborek EDTA with a pH of 8.7. So in general your buffer okay so in general your buffer is very important number one because it maintains your pH and the ionic strength of your system it also allow the flow or the influx of current through your through your medium and most importantly they maintain the pH so that it would not affect okay the buffer would not affect the electrophoretic mobility of your samples or your particles okay so aside from that okay so since we're talking about still we're talking about buffer so remember that here are the possible cases when it comes to any changes or abnormality when it comes to your pH so pH if the pH is acidic okay your hydrogen ion would bind okay your hydrogen ion would bind to your molecule so the ampullitic for example your protein would become positively charged and instead of migrating since they are now positively charged okay since they are now positively charged they will now migrate to your cathode okay so they will now migrate to your cathode so instead of migrating towards the ano okay towards migrating the ano they will migrate the opposite direction so that's a big problem similarly if the pH is also basic or alkaline it would now lose your hydrogen ion so the ampullite now will become negatively charged so in a nutshell what I'm just trying to say here is that any changes on your pH would literally change the electrophoretic mobility of your substance so it's important to maintain your pH okay so pH at 8.6 or 8.7 depending on what is available in your laboratory and what is the optimal buffer that can be used in preserving the integrity of your substances aside from that we'll also have your ionic strength so similarly to your pH any changes on your pH might affect your might affect your might affect your particle in pH it would affect that charge of your particle it would either become a positive or negatively charged ion when it comes to ionic strength on the other hand ionic strength has something to do with your the speed or the the speed of your electrophoresis or the speed of your molecule or your particle so if there is a low ionic strength more charge will be carried so meaning to say more electricity will be carried so the faster the mobility would be so if it is a high if there is high ionic strength on the other hand there will be less charge that will be carried so that would cause a slower mobility so ionic strength is somehow related later on to your electroendosmosis okay your electroendosmosis so remember that when it comes to your buffer it affects your pH and it also affects your ionic strength so if my ionic strength is very low at the mobility okay the mobility of the particle would be faster if the ionic strength is low okay rather if the ionic strength is high rather that there will be less electrical charge that will pass through the medium it would lead to a slower mobility of your particle okay so again the ionic strength ionic strength is inversely proportional to the rate of mobility or the mobility of your the mobility of your particle so when I'm talking about mobility we're talking about the speed the speed on which your particle or your molecule will be passing through your support medium okay so again that is buffer important in maintaining your pH and your ionic strength to maintain the integrity or to maintain the charge of your particle and to maintain the the rate of mobility the rate of mobility within the medium and also let us also not forget forget that your buffer is also important in in allowing the charge or the electrical energy to pass through the medium okay so that is your buffer now after your buffer okay we also have now your support medium or your support media so your support medium these are network of interacting fibers or polymers that is solid but traps large amount of solvent in its pores or channels inside so remember remember that your support media okay your support media these are actually the one that would allow the separation of your components okay they will allow the separation of your component so take for example your your strainer okay your strainer it would look your strainer it is like a mesh or a network of fibers correct or a network of polymers similarly to your support media that is also the idea for it okay so it would it would prevent okay it would prevent bigger molecules to pass through that's why remember when it comes to the factors that affect your electrophoretic mobility the size of the molecule or the particle is a factor when it comes to their mobility so the larger molecules would migrate slower the smaller okay the smaller particle or the smaller molecules would have easier time passing through the support media okay so that's the idea there okay so in that case we're able now to separate the bigger and the larger molecules so always remember that the support media should not interact with the analyte so meaning to say when we say do not interact with your analyte similarly to your PH or rather similarly to your buffer your support media should not affect should not affect the particle so that they will not change their charge they will not change their mobility and they will not impede the movement of molecules through the medium so perfect examples of your support media are your cellulose acetate we can also use your agarose gel and your polyacrylamide gel so these are the three most common support media being used in the laboratory and they're actually specific use for each one of them so we're going to go and discuss about them now okay so when we're talking about your cellulose acetate usually it is best used for isoelectric focusing so your cellulose acetate it's obviously an acetylated cellulose okay to form cellulose acetate by treating it okay your cellulose will treat it with your acetic anhydride to create your cellulose acetate so when you're trying to separate your proteins okay we can separate your protein okay we can separate your cellulose protein into five bands so remember that when we're trying to separate your protein we can separate them by according to their sizes and according to their mobility okay so we have your albumin your alpha one your alpha two blobeins okay alpha one alpha two your beta in your gamma so that is five bands all in total so if we're trying to separate your protein into five fraction okay if you're trying to separate them into five fraction then for example you just want to know where are the alpha alpha one alpha two the beta or the gamma we'll be discussing this in the during the midterm during the semi-final period semi-final period in Glenchem so the different types of protein when we're talking about different types of protein we're not talking about the different amino acids but really different polypeptide or proteins that are inside our body so there if you want to separate your protein you can use your cellulose acetate because that would suffice the need to separate them into five bands Albumin you have your alpha one alpha two globulins globulins you have your beta and you have your gamma globulins okay so those are the five those are the five fractions of your protein so cellulose acetate would be good would be a great support medium for your proteins so aside from your cellulose aside from your cellulose acetate we have here your agarose gel so your agarose gel use it use a purified fraction of agar so the good thing about your agarose gel is it is neutral and thus it does not produce electroendosmosis so what do you mean by electroendosmosis electroendosmosis is a phenomenon where there the medium creates an ionic cloud okay and remember your ionic cloud okay when we have an ionic cloud it can either increase or decrease your ionic strength okay so if there is an increase or decrease in your ionic strength okay the mobility will be affected not only that that ionic cloud can also affect your pH so if your pH is affected not only is the mobility of the particle will be affected their charges will also be affected so in general okay in general your electroendosmosis is the interference in electrophoresis whereby the mobility of your particle is affected okay specifically their pH and their ionic strength okay the pH and the ionic strength within the system okay so if you would want to avoid electroendosmosis you would want to use your agarose gel okay you would want to use your agarose gel so unlike your cellulose actually that it can only divide a particular solution to five fraction your proteins rather it can only divide your proteins into five fraction your agarose gel is good in this in this sense because it can separate your protein into 10 to 15 bands so if you would want to read in advance you can actually scan and you can actually take a look at how many proteins there are in the body there are actually a lot you have your alpha transferin hemopexin your immunoglobulins IGA MEAD so there are actually a lot of proteins inside the body so awal ba if you just simply use your cellulose acetate you can just divide them into five okay and if you divide it into five di mo may identify kung si no yung nandon sa loob ng band again if you have only five bands you cannot specifically point out what specific protein is in there unlike if you use your agarose cell that is a 10 10 to 15 bands you can specifically find out this exact location and then sac density of a particular protein in a particular from a particular solution okay that's the goal when it comes to your in your support media so I don't know if you're getting it but hopefully with this with this analogy you'd be able to somehow get a glimpse of what this support media does okay so remember that when it comes to take for example you're preparing your clothes for laundry isn't it when you when you prepare your clothes for laundry okay when you prepare your clothes for laundry the usual we separate the white we separate the light color from the color to one so take for example just three classification so whether that is a pants a trouser a jacket a t-shirt or a polo okay you will just be separating them according to color okay when you say according to color okay all whites or all light colors or all light color and all dark color shirts or garments or clothes will be grouped together okay that's the thing about cellulose acetate we just group them according to their size or we just group them according to a particular characteristic of your protein but when it comes to sell your clothes cell we're becoming more specific so when we say more specific we're trying not to separate the white shirts the white inner shirts okay the white pants the white polo from the light polo from the light shirt from the dark shirt from the dark pants they're getting my point so with the help of depending on the support media you can you can separate your solution into a more specific component okay so instead of just separating take for example sodium chloride so you can now separate them you can separate the sodium and you can separate the chloride so that's just an example so I hope I hope you're understanding me when it comes to your support media or your support medium being able to separate your protein into different number of bands and what are these bands are for okay so lastly we have your polyacrylamide okay your polyacrylamide separate your protein based on charge and also your molecular charge and also your molecular size okay so when we talk about polyacrylamide it is very much useful in isoenzyme determination so remember I hope everybody still remember the different classes of your enzyme they have your oxidoreductase you have your transferase your hydrolysis your lyases isomerases and also lyases so we have six classes of enzyme so those six classes of enzymes take for example let me just take your lactate dehydrogenase your lactate dehydrogenase they do have five isoenzyme so you have your LDH 1, 2, 3, 4, and 5 so if you want to separate them further if you want to separate them further you might actually want to use your polyacrylamide gel because your polyacrylamide gel is best used for isoenzyme determination and unlike your cellulose acetate it can only divide it to five bands your cellulose acetate it can only divide the bands to 10 to 15 when it polyacrylamide gel can do better it has better resolution it can separate your protein into 20 or more fractions so here you can really point out where does your IGA unlike take for example on the other hand when it comes to your cellulose acetate I'll give an example you have your in the gamma region you want to look into the different immunoglobulins so you would use your cellulose acetate so you can separate now your IGG IGAM END but there are also different isoforms or iso there are also different forms of IGG take for example so you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can you can in isoenzyme determination okay. So again your polyacrylamide gel based its separation through charge and also your molecular side. So in a nutshell we have your support media okay we have your support media so your support media can be a cellulose, acetate, agarose, gel and a polyacrylamide gel okay a polyacrylamide gel so those are the three support media that i will be discussing for this time so with that being said okay this is still your support media so your support media they we do have your in your support media we do have the wells prepared already on your support media so as you can see this is an example of your support media with your wells and this is how we dispense your samples okay this is how we dispense your sample so we have your um this is an automatic pipette so an automatic pipette and we're transferring okay we're transferring the uh we're transferring the substance okay or the sample into the well okay we're transferring it to the well now finally okay so once you dispense your sample on the well um you simply apply the powers worse and then it will automatic it will now run the electrophoresis and will separate your solution into its component into different particles and molecules but the bigger question now sir after separating them how would i know or how would i identify a particular substance isn't it that's what we're after at the end of the day how would i be able to identify if what i actually isolated or what i actually separated here are this this specific protein that i'm looking for and then you know that your electrophoresis can also be used to quantify okay to quantify the substances that you're actually looking for if you won't want to do that okay if you would want to do that you can actually um you can actually um use a densitometer okay if you use a densitometer you would be able to identify the concentration of a particular substance that you um subjected into your electrophoresis so in that case so our detecting system now okay would come into play but before that remember that the result of your electrophoresis consisting now the separated stand of your macromolecules is generally called your electrophoretogram so an example of here on your screen right now these are examples of electrophoretogram so um take for example we have samples one to eight okay samples one to eight these are actually the substances or the samples sir what about the m okay the m there stands for your controls okay so as you can see um with the help of your control you would be able now to identify okay your controls are known patterns already electrophoretic pattern so uh we already know what substance would migrate in this particular region all throughout um until the anode the anode of your um all throughout the anode of your electrophoresis system okay so as you can see here the picture on your the one on your left if you're facing your screen okay the one on your left we have m okay the m are your control and from wells number one to eight those are your those are your samples as you can see you'd be able not to identify okay this is a particular example this is your um alpha two um proteins because they all migrated in this particular band so that would um actually help you there so we also have here on the other hand so here you can now identify okay you can identify the different strands for different substances that are inside your solution okay this is the beauty about electrophoresis okay um this is the beauty about electrophoresis it would allow you now to identify uh a particular substance or a particular particle better okay but just like what i was mentioning a while back um with the use of your electrophoretogram you can actually um you can actually um measure them okay so when it comes to detecting system first okay so when you use kese when you use your electrophoresis it will just be a piece of um support medium so how will you be able to detect what um compound are in there so you can either use your direct observation so if uh direct observation so you can just simply look at the agar or you can simply look at the support medium and you can already identify okay the different um particles or biomolecules that you already have separated you can also use staining so you can use a specific um um you can use a chemical with a specificity um for one chemical group so take for example these are uh no-plique acid so you can use a stain that would that would bind to the phosphate group of your your no-plique acid so you'd be able to identify and you would be able to stain them okay so um you can use your stain aside from that you can also use your radioactive dye so in here you can use your iodine 125 your iodine 125 as a radioactive dye so um disclaimer when it comes to radioactive dye um a big um a big consideration lang because of course um this radio this radioactive dye are um being uh monitored by our pnri so all we remember um to also be cautious in using radioactive um radioactive uh substances and component in the laboratory so aside from direct um observation staining radio using radioactive dye you can actually use your uv visualization so this is the simplest way to detect okay um simplest way to detect your um band so you just simply have a uv light okay you have your uv light um and then you put the support medium on top you'll be able now to um generally you'll be able now to um identify your substances like this one on the photo on your left okay so this is an a uv visualization technique okay so finally ian so you can also so those are observed um direct observation methods okay you can stain them you can use your radioactive dye you can use your uv visualization and if you would want to further identify the concentration of a particular substance in this um in your electrophoresis you can use your densitometer so your densitometer is a device that measures the degree of darkness so the darkness represent the optical density oops if you still remember what optical density is that is the absorbance okay the absorbance so um in this case the the amount of optical density is directly proportional to the concentration of the substance or the component or the particle that we're measuring so the degree of darkness of a photographic or semi-transparent material or of a reflecting um surface so again in densitometer we measure the degree of darkness or the degree of optical that the optical density in other words the absorbance of that molecule so the higher the absorbance or the higher the optical density i'll just use optical density not to um so that you won't be confused here so the the the amount okay the amount or the degree of your optical density is directly proportional to the concentration of the particle or the biomolecule that we're trying to detect using your electrophoresis so in a nutshell that's your electrophoresis very quick and very short discussion so hopefully uh you understand you understood my discussion for today so again please do remember the different components of your electrophoresis the purpose of electrophoresis the factors affecting the mobility the important fact the important consideration when it comes to your buffer their ph or ionic strength your remember what is electro endosmoses this remember the different support medium that if the number of fractions that they can um separate your solution into and of course the different um detecting system um and also your densitometer so with that thank you so much for listening everybody so please do remember i am so this is the i'm on the sky right now so always remember that your attitude um more than your aptitude it will be the one that will determine your altitude so again thank you so much for listening so if you have any questions or clarification please um feel free to send me a message so that i'll be able to guide you and help you understand the topic that we had today so with that thank you so much so this has been um jomar adams gindi so if you have any questions just feel free to send me a message through our tlc through my email or you can simply just message me on messenger so with that thank you so much and have a great day