Abcam, ab205714, STAT 5 A/B ELISA Kit

Size: 1 x 96Tests
STAT 5 A/B ELISA Kit is a single-wash 90-min Simplestep used to quantify STAT 5 A/B 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 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.
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.
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.
Signal Transducer and Activator of Transcription 5 commonly known as STAT5 is a member of the STAT protein family. It weighs approximately 94 kDa. STAT5 usually exists in two closely related forms STAT5a and STAT5b with both playing similar roles but having slightly different activity. This protein is widely expressed found in cells like lymphocytes epithelial cells and endothelial cells indicating its importance in various tissues.
Biological function summary
STAT5 acts as a transcription factor that modulates gene expression by binding to specific DNA sequences. It often forms part of a larger complex when activated. This complex relocation to the nucleus triggers transcriptional activities essential for cell growth differentiation and survival. As it interacts with other proteins STAT5 plays an important role in cytokine signaling particularly in response to interleukins and growth hormones.
Pathways
STAT5 is important in the JAK-STAT signaling pathway. This pathway is activated by cytokines and growth factors leading to STAT5 phosphorylation and its subsequent translocation into the nucleus. The interaction with Janus kinases (JAKs) facilitates this process. STAT5 also plays a significant role in the MAPK pathway linking it to various cellular responses and functions.
STAT5 has a strong association with oncological conditions like leukemia and lymphomas. Aberrant activation or dysregulation of STAT5 can lead to uncontrolled cell proliferation and survival contributing to cancer. Furthermore STAT5 is linked with autoimmune diseases with its malfunction affecting immune response and leading to disorders. In these conditions proteins like BCL-XL often interact with STAT5 influencing cell survival pathways.