STAT5 A/B pY694/699 ELISA Kit (ab176656)
- Datasheet
- References
- Protocols
Overview
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Product nameSTAT5 A/B pY694/699 ELISA Kit
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Detection methodColorimetric
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Precision
Intra-assay Sample n Mean SD CV% A431 extract 6 2.8% Inter-assay Sample n Mean SD CV% A431 extract 3 4% -
Sample typeCell culture extracts, Adherent cells, Suspension cells, Tissue Homogenate
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Assay typeSemi-quantitative
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Sensitivity< 5 µg/ml
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Range10 µg/ml - 1000 µg/ml
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Assay time1h 30m
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Assay durationOne step assay
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Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat
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Product overview
Abcam’s STAT5 A/B (pY694/699) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of STAT5 A/B (pY694/699) protein in Human cells.
The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm
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Notes
Abcam’s STAT5 A/B (pY694/699) SimpleStep ELISA kit (ab176656) has been re-developed with new detector antibody. From February 18, 2016 the kit will be delivered with a new detector total STAT5 antibody. This change was necessary because of a decrease in the performance of the polyclonal STAT5 antibody that was previously used for detection in recent production lots. We have identified a new antibody to use in the SimpleStep ELISA platform that provides higher performance.
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Tested applicationsSuitable for: Sandwich ELISAmore details
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PlatformMicroplate
Properties
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Storage instructionsStore at +4°C. Please refer to protocols.
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Components 1 x 96 tests 10X Wash Buffer PT 1 x 15ml 50X Cell Extraction Enhancer Solution 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 12ml Lyophilized STAT5 A/B Control Lysate 1 vial Plate Seal 1 unit SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit STAT5 A/B (pY694/699) Capture Antibody 1 x 3ml STAT5 A/B (pY694/699) Detector Antibody 1 x 3ml Stop Solution 1 x 12ml TMB Substrate 1 x 12ml -
Research areas
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FunctionCarries out a dual function: signal transduction and activation of transcription. Binds to the GAS element and activates PRL-induced transcription.
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Sequence similaritiesBelongs 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 localizationCytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation.
- Information by UniProt
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Alternative names
- Mammary gland factor
- MGF
- Mpf
see all -
Database links
- Entrez Gene: 6776 Human
- Entrez Gene: 6777 Human
- Entrez Gene: 20850 Mouse
- Entrez Gene: 20851 Mouse
- Entrez Gene: 24918 Rat
- Entrez Gene: 25126 Rat
- Omim: 601511 Human
- Omim: 604260 Human
see all
Applications
Our Abpromise guarantee covers the use of ab176656 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Sandwich ELISA | Use at an assay dependent concentration. |
Images
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Typical cell lysate dilution seriesExample of a typical STAT5 A/B (pY694/699) cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.
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Induction of STAT5 A/B (pY694/699) phosphorylation in A431 cells in response to EGF treatment.A431 cells were cultured in 96-well tissue culture plates and treated (15 min) with a dose-range of EGF before cell lysis. Data from quadruplicate measurements of STAT5 A/B (pY694/699) are plotted and compared against Total STAT5 A/B levels.
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Cell line analysis for Total STAT5 A/BCell line analysis for Total STAT5 A/B from 100 µg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).
Datasheets and documents
Protocols
References
ab176656 has not yet been referenced specifically in any publications.
