Pay attention to the dilution factor for this kit

April 06, 2015

University of Wisconsin-Madison
Department of Food Science
Research Assistant

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Company:

Hycult

Product Name:

EndoCab IgM

Catalog Number:

HK504-IgM

I am interested in the change in EndoCab IgM after 11 weeks of yogurt consumption in humans. My hypothesis is that yogurt consumption can improve intestinal health, therefore reducing trans-location of endotoxin in the gut. EndoCab IgM can be a surrogate marker for circulating endotoxin level. Increased endotoxin exposure tends to deplete the IgM level, thus there is an inverse association between endotoxin level and IgM.

Experimental Design and Results Summary

Application

To determin EndoCab IgM level in human plasma samples

Starting Material

Human EDTA plasma

Protocol Overview

1) Bring samples and reagent to room temperatures. 2) Prepare the dilution buffer and standard stock solution. 3) Dilute plasma samples. 4) Follow the kit insert: load the samples, control and standard---incubate ----add conjugate solution----incubate ----add TMB-----incubate--add stop solution---read

Tips

Make sure the incubation is under 37 degrees, and cover with a lid or adhesive sealing film

Results Summary

The results are pretty consistent within the plate. But I do notice a slightly larger between-batch variance (Could due to the incubation temperature variance).

Additional Notes

None

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Summary

The Good

Working fine. Being used in a lot publications

The Bad

A little pricey compared with other ELISA kits; the dilution of samples should be tested to get reliable results

The Bottom Line

I will continue to buy the kits. Pay attention to the dilution factors because the standard curve is not linear. Difference in ODs could lead to quite different results so figure out the proper dilution factor and stick to it.

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