|Sample type||Average %||Range|
|Cell culture extracts||93.17||81% - 105%|
|Tissue Extracts||92.48||80% - 104%|
Abcam’s TNF alpha ELSA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Rat TNF alpha in cell lysates and tissue lysates.
This assay employs an antibody specific for Rat TNF alpha coated on a 96-well plate. Standards and samples are pipetted into the wells and TNF alpha present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Rat TNF alpha antibody is added. After washing away unbound biotinylated antibody, HRPconjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of TNF alpha bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
|Components||1 x 96 tests|
|200X HRP-Streptavidin Concentrate||1 x 200µl|
|20X Wash Buffer Concentrate||1 x 25ml|
|2X Cell Lysate Buffer||1 x 5ml|
|5X Assay Diluent||1 x 15ml|
|5X Sample Diluent Buffer||1 x 10ml|
|Biotinylated anti-Rat TNF alpha (lyophilized)||2 vials|
|Recombinant Rat TNF alpha Standard||2 vials|
|Stop Solution||1 x 8ml|
|TMB One-Step Substrate Reagent||1 x 12ml|
|TNF alpha Microplate (12 x 8 wells)||1 unit|
Our Abpromise guarantee covers the use of ab100785 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||Use at an assay dependent concentration.|
Rat TNF alpha detected in cell lysates from NR8383.1 control cells (C) or cells stimulated for 6 hours with 5 ug x mL-1 of LPS (Sigma). Results shown for TNF alpha levels in 10e6 cells after background signal was subtracted (duplicates +/- SD).
Rat TNF alpha detected in cell supernatants from NR8383.1 control cells (C) or cells stimulated for 6 hours with 5 ug x mL-1 of LPS (Sigma); background signal subtracted (duplicates +/- SD).
Representative standard curve using ab100785