ELISA Max Kit

December 30, 2013

Overall

Quality of Results

Ease-of-Optimization

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Touro University-CA
Basic Sciences, College of Osteopathic Medicine
Professor

Company:

Biolegend

Product Name:

Human TNFα ELISA MAX™ Deluxe Kit

Catalog Number:

430204

This kit contain all the reagents and supplies needed to perform ELISA on five plates for human IL1b. We tested both serum samples and tissue culture supernatants.

Experimental Design and Results Summary

Application

Sandwich ELISA

Starting Material

Human serum; B lymphoblastoid cell line (BLCL) culture supernatants

Protocol Overview

B lymphoblastoid cell lines (BLCL) were obtained and cultured in OptiMEM medium with 1% fetal calf serum. Viable cells were counted using the Vi-Cell XR Cell Viability Analyzer in order to obtain 3x106 cells. Each sample was cultured in both the presence and absence of 0.05 µg/mL PMA overnight. Cells were then spun down for 5 minutes and supernatants were collected. As a positive control, Thp1 cells were also cultured under the same conditions as the BLCL.

Tips

For washing steps, simply dunk the entire plate into a 2 L bucket containing TBS-T, 3x, and blot dry on a paper towel.

Results Summary

The kit performed outstandingly. Excellent results were obtained from the first time out. The instructions are well written and the reagents labeled clearly. The only reagents needed to be provided by the user will be 2-3 L of TBS-T or PBS-T, and stop buffer.

Additional Notes

At list price, the five-pack kit costs $66/plate. AAI members get an additonal 15% discount. This is well below the cost of a typical ELISA kit, and cost effective compared to "make your own" kits with just standards, capture and detection antibody.

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Summary

The Good

Easy to use, cost-effective, has nearly all reagents needed.

The Bad

Some kits in the series (like the human IL1beta kit) use different sample volumes and standard concentrations than other in the series, so some confusion might occur if running the kits simultaneously. High initial cost outlay compred to strict "roll your own" ELISAs

The Bottom Line

Great kit; I will be purchasing more for future experiments.

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