Human/Mouse STAT5b Antibody
R&D Systems, part of Bio-Techne | Catalog # AF1584
Key Product Details
Validated by
Knockout/Knockdown, Biological Validation
Species Reactivity
Validated:
Human, Mouse
Cited:
Human, Mouse
Applications
Validated:
CyTOF-ready, Immunocytochemistry, Immunoprecipitation, Intracellular Staining by Flow Cytometry, Knockout Validated, Simple Western, Western Blot
Cited:
ChIP (chromatin immunoprecipit, Immunocytochemistry, Immunohistochemistry, Immunoprecipitation, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
Product Specifications
Immunogen
Human STAT5b synthetic peptide
aa 777-787
aa 777-787
Specificity
Detects human and mouse STAT5b.
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Scientific Data Images for Human/Mouse STAT5b Antibody
Detection of Human and Mouse STAT5b by Western Blot.
Western Blot shows lysates of HeLa human cervical epithelial carcinoma cell line, Daudi human Burkitt's lymphoma cell line, NIH-3T3 mouse embryonic fibroblast cell line and M1 mouse myeloid leukemia cell line. PVDF membrane was probed with 0.2 µg/ml of Rabbit Anti-Human/Mouse STAT5b Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1584) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for STAT5b at approximately 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
STAT5b in HepG2 Human Cell Line.
STAT5b was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Rabbit Anti-Human/Mouse STAT5b Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1584) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI(blue). Specific staining was localized to cytoplasm and nucleus. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunoprecipitation of Human and Mouse STAT5b.
HeLa human cervical epithelial carcinoma and M1 mouse myeloid leukemia cell line were untreated (-) or activated (+) with 100 ng/mL Recombinant Human IFN-gamma (285-IF) for 15 minutes. STAT5b was immunoprecipitated from lysates of 5 x 106cells following incubation with 2 µg Rabbit Anti-Human/Mouse STAT5b Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1584) for 1 hour at room temperature. STAT5b-antibody complexes were absorbed using Protein A sepharose (Invitrogen, Catalog # 10-1041). Immunoprecipitated STAT5b was detected by Western blot using 1 µg/mL Human Phospho-STAT5a/b (Y699) Antigen Affinity-purified Polyclonal Antibody (AF4190). View our recommended buffer recipes for immunoprecipitation.
Applications for Human/Mouse STAT5b Antibody
Application
Recommended Usage
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Immunocytochemistry
5-15 µg/mL
Sample: Immersion fixed HepG2 human hepatocellular carcinoma cell line
Sample: Immersion fixed HepG2 human hepatocellular carcinoma cell line
Immunoprecipitation
1-5 µg/500 µg cell lysate
Sample: HeLa human cervical epithelial carcinoma cell line and M1 mouse myeloid leukemia cell line activated with Recombinant Human IFN‑ gamma (Catalog # 285-IF), see our available Western blot detection antibodies. Cell lysate of HAP1 human near-haploid cell line
Sample: HeLa human cervical epithelial carcinoma cell line and M1 mouse myeloid leukemia cell line activated with Recombinant Human IFN‑ gamma (Catalog # 285-IF), see our available Western blot detection antibodies. Cell lysate of HAP1 human near-haploid cell line
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
Sample: Jurkat human acute T cell leukemia cell line fixed with paraformaldehyde and permeabilized with methanol
Sample: Jurkat human acute T cell leukemia cell line fixed with paraformaldehyde and permeabilized with methanol
Knockout Validated
A) STAT5b
is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in
STAT5b knockout HeLa cell line. B) STAT5b is specifically detected in the parental HAP1 cell line, but is not detectable in knockout HAP1 cell line.
Simple Western
10 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and NIH‑3T3 mouse embryonic fibroblast cell line
Sample: HeLa human cervical epithelial carcinoma cell line and NIH‑3T3 mouse embryonic fibroblast cell line
Western Blot
0.2 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line, Daudi human Burkitt's lymphoma cell line, NIH‑3T3 mouse embryonic fibroblast cell line, and M1 mouse myeloid leukemia cell line
Sample: HeLa human cervical epithelial carcinoma cell line, Daudi human Burkitt's lymphoma cell line, NIH‑3T3 mouse embryonic fibroblast cell line, and M1 mouse myeloid leukemia cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Reviewed Applications
Read 3 reviews rated 4.7 using AF1584 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. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
*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: STAT5b
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
signal transducer and activator of transcription 5B, STAT5, transcription factor STAT5B
Gene Symbol
STAT5B
Additional STAT5b Products
Product Documents for Human/Mouse STAT5b Antibody
Product Specific Notices for Human/Mouse STAT5b Antibody
For research use only
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