Detection of TNFa Secretion in Cells

Microbiology and Immunology
Loyola University Chicago
Research Associate

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Company:

BioLegend

Product Name:

Purified anti-human TNFa

Catalog Number:

502802

Info: View Product View Product Specs

Browse Similar Products: TNF-alpha Antibodies

THP1 cells were differentiated into macrophage cells and treated with fraction 10 or control overnight. Cells were stimulated with LPS for 4 hours and then cell culture supernatant was collected for detecting TNFa secretion by ELISA. We found fraction treatment inhibited LPS induced TNFa secretion in macrophages.

Experimental Design and Results Summary

Applications

ELISA

Sample

Cell culture supernatant

Primary Incubation

Serially diluted samples of standard protein were added in a capture antibody coated microplate (1.5 ug/ml)

Blocking Agent

5% nonfat milk in PBST

Secondary Incubation

Biotin alpha-human TNFa was plated and incubated at 37 degrees for 1 hr

Tertiary Incubation

HPR-streptavidin

Detection

TMB

Results Summary

Using the antibody pair, the ELISA generated good signal. We find that a fraction from bacteria secretion suppressed LPS induced TFNa secretion in human macrophages. We usually combine it with detecting antibody and protein standard from BioLegend for detecting TNFa in our experiment. It is sensitive enough to detect TNFa standard protein as low as 1 pg/ml.

Additional Notes

None

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Summary

The Good

The capture antibody is sensitive and easy to use.

The Bad

None

The Bottom Line

We are satisfied with this antibody in combination with TNFa standard protein. We can detect 1pg/ml of the standard. It is sensitive and easy to use.

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