Capture Antibody for ELISA to Test Mouse TNF-alpha

Microbiology and Immunology
Loyola University Chicago
Research Associate

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Company:

BioLegend

Product Name:

Purified anti-mouse TNF-α Antibody

Catalog Number:

510802

Info: View Product View Product Specs

Browse Similar Products: TNF-alpha Antibodies

To test the anti-inflammatory effect of substances from bacteria fractions, we measured the secretion of TNFa in mouse macrophage cell lines after LPS stimulation in the presence or absence of the fractions. We needed to quantify the concentration of the TNFa and bought antibody pair and standard from BioLegend. These reagents work pretty well with high sensitivity and reasonable prices.

Experimental Design and Results Summary

Applications

ELISA

Sample

Supernatant from cell culture

Primary Incubation

1.5 ug/ml capture antibody were coated onto plates overnight and incubated with standard for 1 hr at 37 degrees

Blocking Agent

5% nonfat milk in PBST

Secondary Incubation

0.5ug/ml of 2nd antibody was added in each well and incubated for 1hr at 37 degrees

Tertiary Incubation

HPR-streptavidin was added and incubated for 1 hr at 37 degrees (1 to 10,000)

Detection

TMB was used for developing signal

Results Summary

The ELISA generated good signal that is sensitive enough to detect TNFa standard protein at levels as low as 1 pg/ml. Using these reagents we also found that a fraction from the bacterial secretion suppressed LPS induced TFNa secretion in macrophages. We were able to detect around 20ug/ml in some of our samples, so it is sensitive.

Additional Notes

The ELISA is sensitive as long as you use the antibody with recommended concentration

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Summary

The Good

This product has reasonable price, and is easy to use

The Bad

None

The Bottom Line

This is a very good product that I recommend to use if you need test mouse TNF-alpha by ELISA. I haven't tested it for any other purpose

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