Human/Mouse STAT5b Antibody

R&D Systems | Catalog # AF1584

R&D Systems
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Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Knockout Validated, Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, Simple Western, Immunoprecipitation, CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Immunocytochemistry, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP)

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
View all STAT5b Primary Antibodies »

Product Specifications

Immunogen

Human STAT5b synthetic peptide
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 antibody in HepG2 Human Cell Line by Immunocytochemistry (ICC).

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.

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.

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

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 (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.

Detection of Human and Mouse STAT5b antibody by Simple WesternTM.

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.
Western Blot Shows Human STAT5b Antibody Specificity by Using Knockout Cell Line.

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 (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 knockout HeLa cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western Blot Shows Human STAT5b Specificity Using Knockout Cell Line.

Western blot shows lysates of HAP1 human near-haploid cell line and STAT5b knockout HAP1 cell line (KO). Nitrocellulose 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. A specific band was detected for STAT5b at approximately 85 kDa (as indicated) in the parental HAP1 cell line, but is not detectable in knockout HAP1 cell line. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

Detection of Stat5b by Immunoprecipitation.

Immunoprecipitation was performed on cell lysate of HAP1 human near-haploid cell line using 2.0 μg of Rabbit Anti-Human Stat5b Polyclonal Antibody (Catalog # AF1584) pre-coupled to protein G or protein A beads. Immunoprecipitated Stat5b was detected with a Mouse Anti-Stat5b antibody. The Ponceau stained transfers of each blot are shown. SM=10% starting material; UB=10% unbound fraction; IP=immunoprecipitated. Image, protocol, and testing courtesy of YCharOS Inc. (ycharos.com).

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

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 

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

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

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

Reviewed Applications

Read 3 reviews rated 4.7 using AF1584 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

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.

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.

Calculators

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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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

Entrez Gene IDs

6777 (Human); 20851 (Mouse)

Gene Symbol

STAT5B

Additional STAT5b Products

Product Documents for Human/Mouse STAT5b Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human/Mouse STAT5b Antibody

For research use only

Citations for Human/Mouse STAT5b Antibody

Customer Reviews for Human/Mouse STAT5b Antibody (3)

4.7 out of 5
3 Customer Ratings
5 Stars
67%
4 Stars
33%
3 Stars
0%
2 Stars
0%
1 Stars
0%

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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • Human/Mouse STAT5b Antibody
    Name: Tanya Hundal
    Application: Western Blot
    Sample Tested: BaF3 mouse pro-B cell line
    Species: Mouse
    Verified Customer | Posted 09/13/2021
    BA/F3 cells (vector only and STAT3-JAK2 fusion insert) Experimental condition: Cells treated with STAT siRNAs for 72h (with iL3) Western blot: 40ug protein transferred on nitrocellulose membrane & incubated overnight at 4C using 1:1000 antibody dilutions in TBS-T
    Human/Mouse STAT5b Antibody AF1584
  • Human/Mouse STAT5b Antibody
    Name: Tanya Hundal
    Application: Western Blot
    Sample Tested: TLBR3 human T-cell lymphoma cells
    Species: Human
    Verified Customer | Posted 09/13/2021
    Human/Mouse STAT5b Antibody AF1584
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 22729867
    Species: Human
    Verified Customer | Posted 01/05/2015

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Protocols

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