BioLegend
Biotin anti-human TNFa
502904
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To analyze macrophage activation, 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.
ELISA
Macrophage cell culture supernatant
Serially diluted samples or standard protein were added in a capture antibody coated microplate.
5% nonfat milk in PBST
Biotin anti-human TNFa antibody (1ug/ml) was added in plate and incubated at 37 degrees for 1 hr
HPR-streptavidin
TMB
We found that the fraction from bacteria secretion-suppressed LPS induced TNFa secretion in human macrophages. The capture antibody is sensitive and easy to use. We usually combine capture antibody and protein standard from BioLegend for quantifying TNFa in our experiments. It is sensitive enough to detect TNFa standard protein as low as 1 pg/ml.
None
Using the capture and detection antibody pair from Biolegend, the ELISA generated good signal.
The antibody is sensitive and specific for detection/quantification of human TNFa in cell culture supernatant.