STAT 5 A/B ELISA Kit (ab205714)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 5 µg/ml
- Sample type: Cell culture extracts, Cell Lysate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Human
Overview
-
Product name
STAT 5 A/B ELISA Kit
See all STAT5 kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% A431 6 4.3% Inter-assay Sample n Mean SD CV% A431 3 2.1% -
Sample type
Cell culture extracts, Cell Lysate -
Assay type
Semi-quantitative -
Sensitivity
5 µg/ml -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat
-
Product overview
Abcam’s STAT5 A/B in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit (ab205714) is designed for the semi-quantitative measurement of STAT5 A/B protein in Human cells. STAT5 A and STAT5 B (collectively known as STAT5 A/B) are two very closely related proteins. Although encoded by different genes, STAT5 A and STAT5 B share 96% identity at the protein level. Similarly to other STAT proteins, STAT5 A and STAT5 B are activated by tyrosine phosphorylation, usually by JAK proteins, at Tyr694 and Tyr699, respectively.
An alternative fluorescent substrate, ADHP, can be used with this assay. A microplate reader capable of measuring fluorescence is required if using this product.
As of October 2019, this kit was reformulated with new antibodies to maintain continued long term supply.
-
Notes
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.
Abcam’s STAT5 A/B SimpleStep ELISA kit (ab205714) has been re-developed with new detector antibody. From February 18, 2016 the kit will be delivered with a new detector 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.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
-
Storage instructions
Store at +4°C. Please refer to protocols. -
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 1 x 12ml Lyophilized STAT5 A/B Control Lysate 1 vial Plate Seal 1 unit SimpleStep Pre-Coated 96 Well Microplate (12 x 8 well strips) 1 unit STAT5 A/B (Total) Capture Antibody 1 x 3ml STAT5 A/B (Total) Detector Antibody 1 x 3ml Stop Solution 1 x 12ml TMB Substrate 1 x 12ml -
Research areas
-
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
-
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
- Entrez Gene: 20851 Mouse
- Entrez Gene: 20850 Mouse
- Entrez Gene: 25126 Rat
- Entrez Gene: 24918 Rat
- Omim: 601511 Human
- Omim: 604260 Human
see all
Images
-
Other - STAT 5 A/B ELISA Kit (ab205714)
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
-
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.
-
Cell line analysis for Total STAT5 A/B from 100 µg/mL preparations of cell extractsData from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).
-
Example of a typical STAT5 A/B cell lysate dilution seriesBackground-subtracted data values (mean +/- SD) are graphed.
