Great Multiplex ELISA Product

December 23, 2013

Overall

Quality of Results

Ease-of-Optimization

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Quality Biotech
product validation
research scientist

Company:

Quansys Biosciences

Product Name:

Q-Plex Human Cytokine – Screen (16-plex)

Catalog Number:

#110951HU

Research Aim: Assay validation Experiment Goals: To verify that assays we use in sample testing meet chosen criteria for parameters such as accuracy precision, and linearity of dilution. The particular protocol and results described are from the initial evaluation of linearity of dilution.Reasons for selecting product: higher through-put than classic single elisas

Experimental Design and Results Summary

Application

Bulk sample testing

Starting Material

Commercial human serum or plasma (some individual, some pooled)

Protocol Overview

Samples were received and stored at -80oC. Preparation for the experiment involved installing the free Q-View Software, setting up the Q-View imager using the included set up wizard, preparing the 1X wash buffer from concentrate, and calibrating a plate shaker. Running the kit required a typical chemiluminescent sandwich ELISA protocol: preparing the standard curve (run in duplicate) and samples in the provided diluent, a 1 hour incubation steps for the antigen and then detection antibodies with washes in between, adding strepavadin-conjugated HRP for 15 mins, then substrate. To measure the chemiluminescent signal, the Q-View imager was used as recommended to take 30, 60, and 180 sec exposures. Data was analyzed in the Q-View software using a 5PL regression model.

Tips

The kit manual is long, but quite thorough, with examples and tips

Results Summary

Samples generally had measurable IL-1a, IL-1b, IL-8, IL-12p70, IL-17, IFNg, TNFa, and TNFb. Cytokine levels for the other analytes were generally below the limit of quantification. To evaluate linearity of dilution for the samples, backfits were calculated. The first several dilutions of the samples generally had backfits of 80%-120% until a 1:64 or 1:128 dilution, and then some 50%-140% beyond that.

Additional Notes

Check kit ranges prior to ordering to know whether you want a normal kit or high sensitivity kit

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Summary

The Good

High-throughput, easy to use free analysis software, analysis tips on their website

The Bad

Lots of set up to use a single kit if you don't already do elisas regularly; would be great for repeat users, though

The Bottom Line

I liked this product for quantifying cytokines in serum and plasma. It was similar enough to a normal elisa that the protocol was easy to follow, but it produces more data. The free analysis software Q-View is easy to use.

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