General description
Signal transducer and activator of transcription 5A (UniProt: P42229) is encoded by the STAT5A (also known as STAT5 ) gene (Gene ID: 6776) in human. The protein encoded by this gene is a member of the STAT family of transcription factors. In response to cytokines and growth factors, STAT family members are phosphorylated by the receptor associated kinases, and then form homo or heterodimers that translocate to the cell nucleus where they act as transcription activators. Phosphorylation of STAT5A at tyrosine 694 (Tyr 694) is a critical event in the activation and function of this transcription factor. This phosphorylation is primarily mediated by Janus kinase (JAK) family members, particularly JAK2, which phosphorylates STAT5A upon cytokine receptor engagement. Additionally, Src family kinases have been shown to directly phosphorylate STAT5A at Tyr 694, further enhancing its transcriptional activity and playing a role in erythropoietin-induced signaling. Structurally, STAT5A comprises several functional domains, including a coiled-coil domain for dimerization, a DNA-binding domain, and a transactivation domain. STAT5A is localized predominantly in the cytoplasm and nucleus but has also been detected in mitochondria. Two alternatively spliced transcript variants have been found for this gene. Dysregulation of STAT5A has been implicated in various diseases and activation of this protein in myeloma and lymphoma associated with a TEL/JAK2 gene fusion is independent of cell stimulus and has been shown to be essential for tumorigenesis. Studies suggest that Tyrosine phosphorylation of both STAT5A and STAT5B is necessary for maximal IL-2 signaling and T cell proliferation. This ZooMAb® recombinant monoclonal antibody, generated by our proprietary technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Jian-Xin, Lin., et al. (2024). Nature comm. 15;7372).
ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them.
Immunogen
KLH-conjugated linear peptide corresponding to 12 amino acids surrounding phosphotyrosine 694/699 from the C-terminal half of human STAT5A/B.
Application
Quality Control Testing
Evaluated by Western Blotting in HeLa cell lysate.
Western Blotting Analysis: A 1:10,000 dilution of this antibody detected STAT5A/B phosphorylated at tyrosine 694/699 in lysate from HeLa cells treated with interferon alpha (IFN- ; 2000U/mL, 20 min) but, not in lysate from untreated cells.
Tested Applications
Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected STAT5A/B phosphorylated on tyrosine 694/699 in lysate from MV4-11 cells (MV4-11cells have been reported to have endogenous basal expression of phospho-STAT5A/B).
Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected STAT5A/B phosphorylated at Tyrosine 694/699 in human breast cancer and human colon cancer tissue sections.
Affinity Binding Assay: A representative lot of this antibody bound phospho-STAT5A/B (Tyr694/699) peptide with at least ten thousand-fold (10,000X) higher affinity than with non-specific control peptide.
Peptide Inhibition Assay: Target band detection in lysate from HeLa cells treated with interferon alpha (IFN- ; 2000U/mL, 20 min) was prevented by pre-blocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Biochem/physiol Actions
Clone 1I22 is a ZooMAb® recombinant rabbit monoclonal antibody that specifically detects STAT5A/B phosphorylated at tyrosine 694/699.
Physical form
Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.
Preparation Note
300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.
Other Notes
We are committed to bringing you greener alternative products, which adhere to one or more of the 12 Principles of Green Chemistry. This antibody is preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed, and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.
Legal Information
ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
| biological source | rabbit |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| conjugate | unconjugated |
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| clone | 1I22, recombinant monoclonal |
| description | recombinant, expressed in HEK 293 cells |
| product line | ZooMAb® learn more |
| form | lyophilized |
| mol wt | calculated mol wt 90.65/89.87 kDa,observed mol wt ~90 kDa |
| purified by | using Protein A |
| species reactivity | human |
| species reactivity (predicted by homology) | porcine, rat, mouse |
| packaging | antibody small pack of 25 |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry. |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation |
| sustainability | Greener Alternative Product |
| technique(s) | affinity binding assay: suitable,immunohistochemistry: suitable,inhibition assay: suitable,western blot: suitable |
| isotype | IgG |
| epitope sequence | C-terminal half |
| Protein ID accession no. | NP_001275647 NP_036580 |
| UniProt accession no. | P42229 / P51692 |
| greener alternative category | Aligned |
| shipped in | ambient |
| storage temp. | 2-8°C |
