ERK1/2 (Thr202/Tyr204) In-Cell ELISA Kit (ab126423)
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
ERK1/2 (Thr202/Tyr204) In-Cell ELISA Kit
See all ERK1 + ERK2 kits -
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
ab126423 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 ERK1/2 (Thr202/Tyr204) 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 ERK1/2 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 ERK1/2 (Thr202/Tyr204) 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-ERK1/2 (Thr202/Tyr204) or anti-ERK1/2 is pipetted into the wells and incubated. The wells are washed, and HRP-conjugated anti-mouse IgG 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.
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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-Mouse IgG Concentrate 1 x 10µl Blocking Buffer Concentrate (5X) 1 x 20ml Fixing Solution 1 x 30ml Uncoated 96-well Microplate 1 unit Mouse anti-ERK1/2 Concentrate (1,000X) 1 x 7µl Mouse anti-Phospho-ERK1/2 (Thr202/Tyr204) Concentrate (1,000X) 1 x 7µl Quenching Buffer Concentrate (30x) 1 x 2ml Stop Solution 1 x 14ml TMB One-Step Substrate Reagent 1 x 12ml Wash Buffer A Concentrate (20X) 1 x 30ml Wash Buffer B Concentrate (20X) 1 x 30ml -
Research areas
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Function
Involved in both the initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors such as ELK1. Phosphorylates EIF4EBP1; required for initiation of translation. Phosphorylates microtubule-associated protein 2 (MAP2). Phosphorylates SPZ1 (By similarity). Phosphorylates heat shock factor protein 4 (HSF4) and ARHGEF2.
Acts as a transcriptional repressor. Binds to a [GC]AAA[GC] consensus sequence. Repress the expression of interferon gamma-induced genes. Seems to bind to the promoter of CCL5, DMP1, IFIH1, IFITM1, IRF7, IRF9, LAMP3, OAS1, OAS2, OAS3 and STAT1. Transcriptional activity is independent of kinase activity. -
Sequence similarities
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
Contains 1 protein kinase domain. -
Domain
The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases. -
Post-translational
modificationsDually phosphorylated on Thr-185 and Tyr-187, which activates the enzyme. Dephosphorylated by PTPRJ at Tyr-187. -
Cellular localization
Nucleus. - Information by UniProt
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Alternative names
- ERK 1
- ERK 2
- ERK-1
see all -
Database links
- Entrez Gene: 5595 Human
- Entrez Gene: 5594 Human
- Entrez Gene: 26417 Mouse
- Entrez Gene: 26413 Mouse
- Entrez Gene: 50689 Rat
- Entrez Gene: 116590 Rat
- Omim: 601795 Human
- Omim: 176948 Human
see all
Images
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In-Cell ELISA - ERK1/2 (Thr202/Tyr204) In-Cell ELISA Kit (ab126423)
A431 cells were stimulated by different concentrations of EGF for 10 min at 37°C.
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Western blot - ERK1/2 (Thr202/Tyr204) In-Cell ELISA Kit (ab126423)Western blot analysis of extracts from 100 ng/ml hEGF treated A431 cells. Phospho-ERK1/2 (Thr202/Tyr204) and ERK1/2 antibodies were used in both detection assays.