Phosphotyrosine In-Cell ELISA Kit (ab126431)
Key features and details
- Assay type: Cell-based (qualitative)
- Detection method: Colorimetric
- Sample type: Adherent cells
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Phosphotyrosine In-Cell ELISA Kit -
Detection method
Colorimetric -
Sample type
Adherent cells -
Assay type
Cell-based (qualitative) -
Assay time
5h 10m -
Assay duration
Multiple steps standard assay -
Species reactivity
Reacts with: Mouse, Rat, Human -
Product overview
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.
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Platform
Microplate
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 1 x 96 tests HRP Conjugated Anti-Phosphotyrosine Concentrate (2000X) 1 x 7µl Blocking Buffer Concentrate (5X) 1 x 20ml Fixing Solution 1 x 30ml Quenching Buffer Concentrate (30x) 1 x 2ml Stop Solution 1 x 14ml TMB One-Step Substrate Reagent 1 x 12ml Uncoated 96-well Microplate 1 unit Wash Buffer A Concentrate (20X) 1 x 30ml Wash Buffer B Concentrate (20X) 1 x 30ml -
Research areas
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Relevance
Phosphotyrosine is an amino acid that occurs in endogenous proteins. Tyrosine phosphorylation and dephosphorylation plays a role in cellular signal transduction and possibly in cell growth control and carcinogenesis. -
Alternative names
- L-Phosphotyrosine
- L-Tyrosine-O-phosphate
- O-Phospho-L-tyrosine
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Images
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In-Cell ELISA - Phosphotyrosine In-Cell ELISA Kit (ab126431)
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.