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
Product nameSTAT 5 A/B ELISA Kit
Intra-assay Sample n Mean SD CV% A431 6 4.3% Inter-assay Sample n Mean SD CV% A431 3 2.1%
Sample typeCell culture extracts, Cell Lysate
Assay time1h 30m
Assay durationOne step assay
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat
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.
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.
PlatformPre-coated microplate (12 x 8 well strips)
Storage instructionsStore 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
FunctionCarries out a dual function: signal transduction and activation of transcription. Binds to the GAS element and activates PRL-induced transcription.
Sequence similaritiesBelongs to the transcription factor STAT family.
Contains 1 SH2 domain.
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
- Mammary gland factor
- STAT1 (Tyr701) In-Cell ELISA Kit (ab126426)
- STAT3 (Tyr705) In-Cell ELISA Kit (ab126427)
- STAT4 (Tyr693) In-Cell ELISA Kit (ab126428)
- STAT6 (Tyr641) In-Cell ELISA Kit (ab126430)
- STAT1 (pS727) ELISA Kit (ab126454)
- STAT1 (pY701) ELISA Kit (ab126456)
- STAT1 (pY701) + total STAT1 ELISA Kit (ab126457)
- STAT3 (pY705) ELISA Kit (ab126458)
- STAT3 (pY705) + total STAT3 ELISA Kit (ab126459)
- STAT3 pY705 ELISA Kit (ab176654)
- STAT3 ELISA Kit (ab176655)
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.
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.
Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).
Background-subtracted data values (mean +/- SD) are graphed.
ab205714 has been referenced in 1 publication.
- Wang T et al. GDF15 is a heart-derived hormone that regulates body growth. EMBO Mol Med 9:1150-1164 (2017). PubMed: 28572090