Species Reactivity
Human, Mouse
Specificity
Detects endogenous human and mouse STAT5A and STAT5B in Western blots.
Source
Polyclonal Rabbit IgG
Purification
Antigen Affinity-purified
Immunogen
Human STAT5a and STAT5b synthetic peptide
aa 219-390Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated
Applications
Recommended
ConcentrationSample
Western Blot
1 µg/mL
See below
Simple Western
20 µg/mL
See below
Chromatin Immunoprecipitation (ChIP)
5 µg/5 x 10 6 cells
See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. are available in the Technical Information section on our website.
Data Examples
Western Blot | Detection of Human/Mouse STAT5a/b by Western Blot. Western blot shows lysates of K562 human chronic myelogenous leukemia cell line and M1 mouse myeloid leukemia cell line. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human/Mouse STAT5a/b Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2168) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # ). Specific bands were detected for STAT5a at approximately 94 kDa and STAT5b at approximately 92 kDa (as indicated). This experiment was conducted under reducing conditions and using . |
Chromatin Immunoprecipitation (ChIP) | Detection of STAT5a/b-regulated Genes by Chromatin Immunoprecipitation. BaF3 mouse pro-B cell line treated with 10 ng/mL Recombinant Mouse IL‑3 (Catalog # ) for 1 hour was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. STAT5a/b/DNA complexes were immunoprecipitated using 5 μg Rabbit Anti-Human/Mouse STAT5a/b Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2168) or control antibody (Catalog # ) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Rabbit IgG Secondary Antibody (Catalog # ). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # ) and DNA was purified using chelating resin solution. The bcl-x promoter was detected by standard PCR. |
Simple Western | Detection of Human STAT5a/b by Simple WesternTM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and K562 human chronic myelogenous leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for STAT5a/b at approximately 91 kDa (as indicated) using 20 µg/mL of Rabbit Anti-Human/Mouse STAT5a/b Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2168). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. |
Preparation and Storage
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: STAT5a/b
Signal transduction and activator of transcription 5 (STAT5) is a member of the Jak/STAT signal transduction pathway and is activated by a variety of cytokines (IL22, IL6, IFN-α). STAT5 has two isoforms (A and B) that share 93% amino acid identity and bind the DNA consensus site TTCN3GAA. STAT5 mediates cytokine signaling by acting as a signal transducer in the cytoplasm and, upon phosphorylation, translocates to the nucleus and activates transcription of specific genes. STAT5 is involved in a wide array of biological processes ranging from regulating apoptosis to adult mammary gland proliferation, differentiation and survival.
Long Name:
Signal Transducer and Activator of Transcription 5b
Alternate Names:
MGF; STAT5; STAT5A signal transducer and activator of transcription 5A ; STAT5a/b
