How to perform a 7 minute protein transfer with the iBlot® - Western Blotting System





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Published on Apr 4, 2012

Please visit: http://ow.ly/9Tpv3 to learn more about western blotting systems.
Please also refer to the How To video http://www.youtube.com/watch?v=g7DT5x... to see how to set up and perform western blot protein transfer (dry protein blotting) using the iBlot®2 Dry Blotting System.

In this video life Technologies staff scientist, David Bourdon Ph.D walks through the steps of using the iBlot® - Western Blotting System to preform a dry western blot.

Video transcription:
How To Perform a 7 Minute Protein Transfer With The iBlot®
Western Blotting System
Life Technologies Corp.

Presented by David Bourdon Ph.D, Life Technologies Staff Scientist

Ok so let's go through the steps of an iBlot. So we have our iBlot apparatus here and let's go ahead and unwrap our anode stack. Put that on the device. And we've got our gel that's ready to go here. Our nupage gel that runs on purified GFP and Novex sharp pre-stained ladder. Ok and we're going to pre-wet our one filter paper, put that on top. Then use our roller to roll out any bubbles.

Now we're going to go ahead and put our cathode stack on top. And then finish of with our sponge on top.

Now we have seven minutes to wait.

Ok, that sound is the completion of our seven-minute iBlot transfer. Let's go ahead and take down the iBlot stack. So first of all, open up the apparatus, take off the sponge, take off the top electrode. Take off the filter paper. Take off our nupage gel. And here is our PVDF membrane. And behold, our proteins of interest, at least we can say for now, the dye fronts transferred as well as the sharp pre-stained standards. So now this blot is ready to put in block, followed by primary antibody wash, secondary-body wash, and follow up with detection. And that's it.

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