Abcam, ab205715, STAT 5 A/B (pY694/699 + Total) ELISA Kit

Size: 1 x 96Tests
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,
Reacts with:Human,
Assay type:Semi-quantitative,
Sensitivity:= 5 µg/mL,
Assay time:1h 30m,
Assay Platform:Pre-coated microplate (12 x 8 well strips)

Product details:
Abcam's STAT5 A/B (pY694/699) and STAT5 A/B (Total) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit (
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 (
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.
REACH authorisation
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.

Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°C

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
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.
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.