BioLegend
Purified anti-mouse TNF-α Antibody
510802
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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.
ELISA
Supernatant from cell culture
1.5 ug/ml capture antibody were coated onto plates overnight and incubated with standard for 1 hr at 37 degrees
5% nonfat milk in PBST
0.5ug/ml of 2nd antibody was added in each well and incubated for 1hr at 37 degrees
HPR-streptavidin was added and incubated for 1 hr at 37 degrees (1 to 10,000)
TMB was used for developing signal
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.
The ELISA is sensitive as long as you use the antibody with recommended concentration
This product has reasonable price, and is easy to use
None
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