For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome
ab126428 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 STAT4 (Tyr693) phosphorylation and screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators in cultured human and mouse cell lines. By determining STAT4 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 STAT4 (Tyr693) 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-STAT4 (Tyr693) or Anti-STAT4 (primary antibody) is pipetted into the wells and incubated. The wells are washed, and HRP-conjugated anti-Mouse IgG or HRP-conjugated anti-rabbit 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|
|5X Blocking Buffer Concentrate (Item F)||1 x 20ml|
|Fixing Solution (Item D)||1 x 30ml|
|Anti-Mouse IgG Concentrate (Item I)||1 x 10µl|
|Anti-Rabbit IgG Concentrate (Item I-1)||1 x 10µl|
|Uncoated 96-well Microplate (Item A)||1 unit|
|Mouse Anti-STAT4 Concentrate (Item H)||1 x 7µl|
|Quenching Buffer Concentrate (30x) (Item E)||1 x 2ml|
|Rabbit Anti-Phospho-STAT4 (Tyr693) Concentrate (Item G)||1 x 10µl|
|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 ab126428 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.|
A431 cells were stimulated by different concentrations of recombinant human EGF for 10 minutes at 37°C
Western blot analysis of extracts from 100 ng/ml hEGF treated A431 cells. Phospho-Stat4 (Tyr693) and Stat4 antibodies were used in both detection assays.
ab126428 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"