Human/Mouse STAT5a/b Pan Specific Antibody
R&D Systems, part of Bio-Techne | Catalog # AF2168
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human, Mouse
Cited:
Human, Mouse
Applications
Validated:
Chromatin Immunoprecipitation (ChIP), Simple Western, Western Blot
Cited:
Chromatin Immunoprecipitation (ChIP), Flow Cytometry, Functional Assay, Neutralization, Simple Western, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
Product Specifications
Immunogen
Human STAT5a and STAT5b synthetic peptide
aa 219-390
aa 219-390
Specificity
Detects endogenous human and mouse STAT5A and STAT5B in Western blots.
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Scientific Data Images for Human/Mouse STAT5a/b Pan Specific Antibody
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 # HAF008). 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 Immunoblot Buffer Group 1.
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 # 403-ML) 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 # AB-105-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Rabbit IgG Secondary Antibody (Catalog # BAF008). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. Thebcl-xpromoter was detected by standard PCR.
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.Applications for Human/Mouse STAT5a/b Pan Specific Antibody
Application
Recommended Usage
Chromatin Immunoprecipitation (ChIP)
5 µg/5 x 106 cells
Sample: Recombinant Mouse IL‑3 (Catalog # 403-ML) treated BaF3 mouse pro-B cell line chromatin, bcl-x promoter detected by standard PCR
Sample: Recombinant Mouse IL‑3 (Catalog # 403-ML) treated BaF3 mouse pro-B cell line chromatin, bcl-x promoter detected by standard PCR
Simple Western
20 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and K562 human chronic myelogenous leukemia cell line
Sample: HeLa human cervical epithelial carcinoma cell line and K562 human chronic myelogenous leukemia cell line
Western Blot
1 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line and M1 mouse myeloid leukemia cell line
Sample: K562 human chronic myelogenous leukemia cell line and M1 mouse myeloid leukemia cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Reviewed Applications
Read 1 review rated 4 using AF2168 in the following applications:
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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
Additional STAT5a/b Products
Product Documents for Human/Mouse STAT5a/b Pan Specific Antibody
Product Specific Notices for Human/Mouse STAT5a/b Pan Specific Antibody
For research use only
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