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Abcam’s Human CD86 in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of CD86 in Human serum, plasma, buffered solutions or cell culture medium.
A monoclonal antibody specific for CD86 has been coated onto the wells of the microtiter strips provided. Samples, including standards of known CD86 concentrations, control specimens or unknowns are pipetted into these wells. During the first incubation, the standards or samples and a biotinylated monoclonal antibody specific for CD86 are simultaneously incubated. After washing, the enzyme Streptavidin-HRP, that binds the biotinylated antibody is added, incubated and washed. A TMB substrate solution is added which acts on the bound enzyme to induce a colored reaction product. The intensity of this colored product is directly proportional to the concentration of CD86 present in the samples.
This kit will recognize both endogenous and recombinant Human CD86.
|Components||Identifier||2 x 96 tests||1 x 96 tests|
|10X Standard Diluent Buffer||Black||1 x 25ml||1 x 25ml|
|200X Wash Buffer||White||2 x 10ml||1 x 10ml|
|Biotinylated Antibody Diluent||Red||1 x 13ml||1 x 7ml|
|Biotinylated anti-CD86||Red||2 x 0.4ml||1 x 0.4ml|
|CD86 Microplate (12 x 8 well strips)||2 units||1 unit|
|CD86 Standard (Lyophilized)||Yellow||4 vials||2 vials|
|Chromogen TMB Substrate Solution||1 x 24ml||1 x 11ml|
|HRP Diluent||Red||1 x 23ml||1 x 23ml|
|Stop Reagent||Black||2 x 11ml||1 x 11ml|
|Streptavidin-HRP||4 x 5µl||2 x 5µl|
Our Abpromise guarantee covers the use of ab45921 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.|
CD86 in U937 lysates from control cells or cells stimulated for 24 hours with 50 ng x mL-1 PMA (ab120297) or PMA with 1 ug x mL-1 LPS for the last 6 hours (duplicates; +/- SD).
2e6 snap frozen cells were lysed in 0.3 mL RIPA lysis buffer on ice, sonicated in an ice cold water bath for 12 intervals for a total of 3.5 minutes. The supernatants were cleared by centrifugation (max speed, 10 minutes, 4 degrees), and tested undiluted or diluted 1/4.
CD86 measured in undiluted 72 hours culture supernatants from control of treated (5 ug x mL-1 PHA; Sigma) PBMCs (1e6 cells x mL-1; duplicates +/- SD).
Standard curve with background signal subtracted (duplicates; +/- SD).
Representative Standard Curve using ab45921
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"