Human STAT5 (Tyr694) In-Cell ELISA Kit (ab126429)
Key features and details
- Assay type: Semi-quantitative
- Detection method: Colorimetric
- Sample type: Adherent cells
- Reacts with: Human
Overview
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Product name
Human STAT5 (Tyr694) In-Cell ELISA Kit
See all STAT5 kits -
Detection method
Colorimetric -
Sample type
Adherent cells -
Assay type
Semi-quantitative -
Assay time
5h 10m -
Assay duration
Multiple steps standard assay -
Species reactivity
Reacts with: Human -
Product overview
ab126429 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 STAT5 (Tyr694) phosphorylation and screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators in cultured human cell lines. By determining STAT5 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 In-Cell STAT5 (Tyr694) 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-STAT5 (Tyr694) or Anti-STAT5 (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.
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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-Phospho-STAT5 (Tyr694) Concentrate (Item G) 1 x 7µl Mouse Anti-STAT5 Concentrate (Item H) 1 x 10µl 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
Carries out a dual function: signal transduction and activation of transcription. Binds to the GAS element and activates PRL-induced transcription. -
Sequence similarities
Belongs to the transcription factor STAT family.
Contains 1 SH2 domain. -
Post-translational
modificationsTyrosine phosphorylated in response to IL-2, IL-3, IL-7, IL-15, GM-CSF, growth hormone, prolactin, erythropoietin and thrombopoietin. Tyrosine phosphorylation is required for DNA-binding activity and dimerization. Serine phosphorylation is also required for maximal transcriptional activity. -
Cellular localization
Cytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation. - Information by UniProt
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Alternative names
- MGF
- Signal transducer and activator of transcription 5A
- Signal Transducer and Activator of Transcription 5B
see all -
Database links
- Entrez Gene: 6777 Human
- Entrez Gene: 6776 Human
- Omim: 601511 Human
- Omim: 604260 Human
- SwissProt: P51692 Human
- SwissProt: P42229 Human
- Unigene: 437058 Human
Images
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In-Cell ELISA - STAT5 (Tyr694) Human In-Cell ELISA Kit (ab126429)
A431 cells were stimulated by different concentrations of EGF for 10 minutes at 37°C.
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In-Cell ELISA - STAT5 (Tyr694) Human In-Cell ELISA Kit (ab126429)A431 cells were stimulated by different concentrations of EGF for 30 minutes at 37°C.
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Western blot - STAT5 (Tyr694) Human In-Cell ELISA Kit (ab126429)Western blot analysis of extracts from 100 ng/ml hEGF treated A431 cells. Phospho-STAT5 (Tyr694) and STAT5 antibodies were used in both detection assays.