STAT 5 A/B (pY694/699 + Total) ELISA Kit is a single-wash 90-min Simplestep used to quantify STAT 5 A/B (pY694/699 + Total) with a sensitivity of 5 µg/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
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- Communications biology 4:7032021Random errors in protein synthesis activate an age-dependent program of muscle atrophy in mice.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJames Moore,Rashid Akbergenov,Martina Nigri,Patricia Isnard-Petit,Amandine Grimm,Petra Seebeck,Lisa Restelli,Stephan Frank,Anne Eckert,Kader Thiam,David P Wolfer,Dimitri Shcherbakov,Erik C BöttgerPubMed 34103648
- Journal of dairy science 101:2588-26012017Effect of 17β-estradiol on milk production, hormone secretion, and mammary gland gene expression in dairy cows.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJ J Tong,I M Thompson,X Zhao,P LacassePubMed 29274969
Key facts
- Detection method
- Colorimetric
- Sample types
- Cell culture extracts, Cell Lysate
- Assay type
- Semi-quantitative
- Reactive species
- Human
- Assay time
- 1h 30m
- Sensitivity
- = 5 µg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
What's included?
Recommended products
STAT 5 A/B (pY694/699 + Total) ELISA Kit is a single-wash 90-min Simplestep used to quantify STAT 5 A/B (pY694/699 + Total) with a sensitivity of 5 µg/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
Key facts
- Detection method
- Colorimetric
- Sample types
- Cell culture extracts, Cell Lysate
- Assay type
- Semi-quantitative
- Reactive species
- Human
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 5 µg/mL
Precision
Intra assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Overall | n 6 | mean - | SD - | C.V. 3.9 |
Sample pY694/699 | n 6 | mean - | SD - | C.V. 2.8 |
Inter assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Overall | n 3 | mean - | SD - | C.V. 2.1 |
Sample pY694/699 | n 3 | mean - | SD - | C.V. 4.3 |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
Abcam's STAT5 A/B (pY694/699) and STAT5 A/B (Total) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit (Tyrosinase Inhibitor Assay Kit (Colorimetric) ab204715) is designed for the semi-quantitative measurement of STAT5 A/B (pY694/699) and Total STAT5 A/B 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
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.
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.
Abcam's STAT5 A/B (pY694/699) and STAT5 A/B (Total) SimpleStep ELISA® kit (Tyrosinase Inhibitor Assay Kit (Colorimetric) ab204715) has been re-developed with new detection antibody. From February 18, 2016 the kit will be delivered with a new detection total STAT5 antibody. This change was necessary because of a decrease in the performance of the polyclonal STAT5 antibody that was previously used for capture 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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
STAT5a and STAT5b collectively known as STAT5 (Signal Transducer and Activator of Transcription 5) are transcription factors involved in signal transduction and activation of transcription. These proteins have a molecular mass of approximately 90 kDa each. They express in many cell types including lymphocytes and hepatocytes. STAT5a and STAT5b share high structural similarity but have distinct functional roles in different tissues. They remain inactive in the cytoplasm until cytokine or growth factor stimulation triggers their phosphorylation enabling them to dimerize and translocate to the nucleus to affect gene expression.
- Biological function summary
STAT5 proteins play an important role in cell growth differentiation and survival. They are not only essential for normal immune function but also form part of a complex network that regulates the expression of specific genes. Dimerization is important for their activation and they can also form heterodimers with other STAT family members. Their activity affects numerous cellular processes from proliferation to apoptosis therefore impacting various physiological functions particularly in hematopoietic and immune systems.
- Pathways
STAT5a and STAT5b involvement is mainly in the JAK-STAT signaling pathway and the growth hormone signaling pathway. Activation of these pathways occurs through interaction with cytokine receptors such as IL-2 and IL-7 which leads to JAK kinases' activation. The activated JAK-STAT pathway plays an essential role in immune regulation and hematopoiesis. STAT5 also interacts with proteins like JAK2 and other STAT family members to propagate signaling cascades that drive cell proliferation and differentiation.
- Associated diseases and disorders
The dysregulation of STAT5 activity links to conditions such as leukemia and breast cancer. STAT5a and STAT5b display upregulated expression or activity in these conditions contributing to uncontrolled cell growth and survival. Their interplay with oncogenic proteins like BCR-ABL in chronic myeloid leukemia highlights their role in tumorigenesis and disease progression. Alterations in STAT5 signaling may also affect interactions with other proteins involved in cancer pathways influencing treatment outcomes and resistance.
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3 product images
Sandwich ELISA - STAT 5 A/B (pY694/699 + Total) ELISA Kit (ab205715)
Cell line analysis for Total STAT5 A/B from 100 µg/mL preparations of cell extracts.
Cell 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).
Sandwich ELISA - STAT 5 A/B (pY694/699 + Total) ELISA Kit (ab205715)
Induction of STAT5 A/B (pY694/699) phosphorylation in A431 cells in response to EGF treatment.
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.
Sandwich ELISA - STAT 5 A/B (pY694/699 + Total) ELISA Kit (ab205715)
Example of a typical STAT5 A/B (pY694/699) and STAT5 A/B (Total) cell lysate dilution series. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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