Human/Mouse STAT5b Antibody

Catalog # Availability Size / Price Qty
AF1584
AF1584-SP
Detection of Human, Mouse, and Rat STAT5b by Western Blot.
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Product Details
Citations (3)
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Human/Mouse STAT5b Antibody Summary

Species Reactivity
Human, Mouse
Specificity
Detects human and mouse STAT5b.
Source
Polyclonal Rabbit IgG
Purification
Antigen Affinity-purified
Immunogen
Human STAT5b synthetic peptide
aa 777-787
Formulation
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 Concentration
Sample
Western Blot
0.2 µg/mL
See below
Simple Western
10 µg/mL
See below
Immunoprecipitation
10 µg/106 cells
See below
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
See below
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
See below
Knockout Validated
STAT5b is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in STAT5b knockout HeLa cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Western Blot Detection of Human, Mouse, and Rat STAT5b by Western Blot. View Larger

Detection of Human, Mouse, and Rat 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, M1 mouse myeloid leukemia cell line, and PC‑12 rat adrenal pheochromocytoma 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 Immunoblot Buffer Group 1.

Immunocytochemistry STAT5b in HepG2 Human Cell Line. View Larger

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; Catalog # 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 Immunoprecipitation of Human and Mouse STAT5b. View Larger

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 (Catalog # 285-IF) for 15 minutes. STAT5b was immunoprecipitated from lysates of 5 x 106 cells 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 (Catalog # AF4190). View our recommended buffer recipes for immunoprecipitation.

Intracellular Staining by Flow Cytometry Detection of STAT5b in Jurkat Human Cell Line by Flow Cytometry. View Larger

Detection of STAT5b in Jurkat Human Cell Line by Flow Cytometry.

Jurkat human acute T cell leukemia cell line was stained with Rabbit Anti-Human/Mouse STAT5b Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF1584, filled histogram) or control antibody (Catalog # AB-105-C, open histogram), followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.

Simple Western Detection of Human and Mouse STAT5b by Simple WesternTM. View Larger

Detection of Human and Mouse STAT5b by Simple WesternTM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and NIH‑3T3 mouse embryonic fibroblast cell line, loaded at 0.2 mg/mL. A specific band was detected for STAT5b at approximately 92 kDa (as indicated) using 10 µg/mL of Rabbit Anti-Human/Mouse STAT5b Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1584). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Knockout Validated Western  Blot Shows Human STAT5b Specificity by Using Knockout Cell Line. View Larger

Western Blot Shows Human STAT5b Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and STAT5b knockout HeLa cell line (KO). 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) in the parental HeLa cell line, but is not detectable in knockoutHeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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: 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
Entrez Gene IDs
6777 (Human); 20851 (Mouse)
Alternate Names
signal transducer and activator of transcription 5B; STAT5; STAT5b; transcription factor STAT5B

Product Datasheets

Citations for Human/Mouse STAT5b Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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  1. Critical functions for STAT5 tetramers in the maturation and survival of natural killer cells
    Authors: JX Lin, N Du, P Li, M Kazemian, T Gebregiorg, R Spolski, WJ Leonard
    Nat Commun, 2017;8(1):1320.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Immunoprecipitation
  2. BCR-ABL affects STAT5A and STAT5B differentially.
    Authors: Schaller-Schonitz M, Barzan D, Williamson A, Griffiths J, Dallmann I, Battmer K, Ganser A, Whetton A, Scherr M, Eder M
    PLoS ONE, 2014;9(5):e97243.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Immunoprecipitation
  3. Rac1 and a GTPase-activating protein, MgcRacGAP, are required for nuclear translocation of STAT transcription factors.
    Authors: Bao YC, Nomura Y, Moon Y, Tonozuka Y, Minoshima Y, Hatori T, Tsuchiya A, Kiyono M, Nosaka T, Nakajima H
    J. Cell Biol., 2006;175(6):937-46.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC

FAQs

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Cell and Tissue Staining Kits

ExactaChIP Chromatin Immunoprecipitation Kits

Isotype Controls

Normal Rabbit IgG Control

Ctrl,CyTOF-ready

Reconstitution Buffers

Secondary Antibodies

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