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ab126425 is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cells. It can be used for measuring the relative amount of p38 MAPK (Thr180/Tyr182) phosphorylation and screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators in cultured human, mouse and rat cell lines. By determining p38 MAPK protein phosphorylation in your experimental model system, you can verify pathway activation in your cell lines without spending excess time and effort in preparing cell lysate and performing an analysis of Western Blot.
In the p38 MAPK (Thr180/Tyr182) In-Cell ELISA Kit, cells are seeded into a 96 well tissue culture plate. The cells are fixed after various treatments, inhibitors or activators. After blocking, Anti-Phospho-p38 MAPK (Thr180/Tyr182) or Anti-p38 MAPK (primary antibody) is pipetted into the wells and incubated. The wells are washed, and HRP-conjugated anti-mouse IgG (secondary antibody) is added to the wells. The wells are washed again, a TMB substrate solution is added to the wells and color develops in proportion to the amount of protein. 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|
|Anti-Mouse IgG Concentrate (Item I)||1 x 10µl|
|5X Blocking Buffer Concentrate (Item F)||1 x 20ml|
|Fixing Solution (Item D)||1 x 30ml|
|Uncoated 96-well Microplate (Item A)||1 unit|
|Mouse Anti-p38 MAPK Concentrate (Item H)||1 x 7µl|
|Mouse Anti-Phospho-p38 MAPK (Thr180/Tyr182) Concentrate (Item G)||1 x 7µl|
|Quenching Buffer Concentrate (30x) (Item E)||1 x 2ml|
|Stop Solution||1 x 14ml|
|TMB One-Step Substrate Reagent||1 x 12ml|
|20X Wash Buffer A Concentrate (Item B)||1 x 30ml|
|20X Wash Buffer B Concentrate (Item C)||1 x 30ml|
Our Abpromise guarantee covers the use of ab126425 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|In-Cell ELISA||Use at an assay dependent concentration.|
HeLa cells were stimulated by different concentrations of anisomycin for 15 minutes at 37°C.
HeLa cells were stimulated by different concentrations of anisomycin for 1 hour at 37°C.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"