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ab126431 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 Phosphotyrosine 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 Phosphotyrosine protein 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 Cell-Based Phosphotyrosine ELISA kit, cells are seeded into a 96 well tissue culture plate. The cells are fixed after various treatments, inhibitors or activators. After blocking, HRP-Anti- Phosphotyrosine is pipetted into the wells and incubated. The wells are washed, 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-Phosphotyrosine-HRP Concentrate (Item E-1)||1 x 7µl|
|5X Blocking Buffer Concentrate (Item F)||1 x 15ml|
|Fixing Solution (Item D)||1 x 30ml|
|Uncoated 96-well Microplate (Item A)||1 unit|
|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 25ml|
|20X Wash Buffer B Concentrate (Item C)||1 x 25ml|
Our Abpromise guarantee covers the use of ab126431 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 treated for 30 min with 50 µL of 100 mM Genistein or 2 mM Lavendustin in appropriate wells at room temperature prior to EGF stimulation. Added 50 µL different concentrations of rhEGF (0, 20 or 100 ng/ml in serum free DMEM) to appropriate wells. Then incubated for 10 min at 37°C.
ab126431 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"