|Sample type||Average %||Range|
|Cell culture extracts||94.53||84% - 104%|
|Tissue Extracts||93.39||83% - 103%|
Abcam’s IL-6 Mouse ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse IL-6 in cell lysates and tissue lysates.
This assay employs an antibody specific for mouse IL-6 coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-6 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse IL-6 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated 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 IL-6 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-mouse IL-6||2 vials|
|IL-6 Microplate (12 x 8 wells)||1 unit|
|Recombinant mouse IL-6 Standard||2 vials|
|Stop Solution||1 x 8ml|
|TMB One-Step Substrate Reagent||1 x 12ml|
Our Abpromise guarantee covers the use of ab100713 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.|
IL-6 measured in mouse tissue lysates prepared with RIPA buffer and sonicated in an ice-cold water bath for 12 intervals of a total of 3.5 minutes. 0.02-1 mg protein x mL-1 were tested, results shown as IL-6 per mg of extracted protein (duplicates +/- SD).
IL-6 measured in RIPA lysates from control cells, or cells stimulated for 24 hours with 50 ng x mL-1 of PMA (ab120297), with the addition of 1 ug x mL-1 of LPS (Sigma) (P/L) for the last 6 hours. Samples were diluted and tested in the range of 1e4-5e5 cells x mL-1, results expressed per million cells; duplicates +/- SD).
Dilution curves of mouse IL-6 (ab100713) and NIBSC standard (93/730). One pg of standard mouse IL-6 corresponds to 0.37 (+/- 0.07) AU. Background signal subtracted (duplicates; +/- SD).
Standard curve of mouse IL-6 with background signal subtracted (duplicates; +/- SD).
Representative standard curve using ab100713