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MAGPIX® System - How To Analyze Protein Data

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Published on Oct 12, 2012

Learn more about MAGPIX® System at http://www.lifetechnologies.com/magpix. The move from multiple protein assays to multiplex assays is now totally within reach!
The MAGPIX® system is an affordable, compact, fluorescent-based detection system based on Luminex's proven xMAP® technology. With the MAGPIX system every researcher can perform quantitative analysis of up to 50 proteins simultaneously using only 25μl of precious sample. The MAGPIX® system is personal, benchtop instrument with out of the box set-up with step by step guide to interactive software and all designed for use with broad and expanding menu of assay kits. For a demonstration of the MAGPIX System, click here!

To qualify your raw data including the standard curve, you'll need to properly analyze the quantitative assay using xPONENT® software. Once you've completed a batch, go to the "Saved Batches" under the Results tab. Next, select the assay that you want to analyze from the list. Open the batch by clicking the "Open" button at the bottom right of the screen.

Before you begin to qualify the standard curves for each analyte make sure that you check the raw data to assure accurate data acquisition. Review the bead scatter plot for any abnormalities such as aggregation, bead shift, air bubbles, and insufficient count. Next, review the plate map while looking for any errors. A successful well will show the color green.

To start this analysis, choose an analyte from the "Analyte" drop-down box in the header. Using a visual inspection of the standard curve, you will want to identify any severe outlier potential plateau, or bottoming-out points. Next, locate the standard points at the top and bottom of the standard curve. It is important to review key statistics such as Net-MFI and % recovery. %Recovery is defined as the observed concentration versus expected concentration. The average range for each standard point should be between 80-120%, which will assure the most accurate results. Any standard points that exceed this range should be invalidated. Repeat analysis for each target.

Here are some examples that might warrant invalidating data points. Yellow warnings in wells indicate possible low bead count due to clogging or aggregation. To confirm, select "Count" from the Statistic drop-down box and review the counts for each target. The recommended target for counts is 100 beads, so any result of less than 100 should be analyzed carefully. The fewer beads counted per analyte, the less accurate and reproducible the result. Counts below 35 are always recommended to be invalidated. To invalidate a sample well, select the target or well and click on invalidate. Then save.

When reviewing the standard curve, if there are data points that exceed acceptable limits such as % recovery, they should be reviewed carefully and possibly invalidated. To invalidate a standard point of one target, highlight that point, click on invalidate, analyze, and then save.

For more information, visit http://www.lifetechnologies.com/magpix

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