Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Simple Western, Chromatin Immunoprecipitation (ChIP)

Cited:

Western Blot, Neutralization, Flow Cytometry, Simple Western, Chromatin Immunoprecipitation (ChIP), Functional Assay

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
View all STAT5a/b Primary Antibodies »

Product Specifications

Immunogen

Human STAT5a and STAT5b synthetic peptide
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 antibody by 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 # 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 antibody by Chromatin Immunoprecipitation.

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 antibody by Simple WesternTM.

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.
Detection of Human STAT5a/b by Western Blot

Detection of Human STAT5a/b by Western Blot

Effects of hypoxia generated by using the insert-based enzymatic system on the phosphorylation of cell signalling molecules. Several molecules associated with cellular signalling and survival events are differentially phosphorylated between the hypoxia (H) and normoxia (N) groups at 2 hours after the hypoxic insult. One representative western blotting experiment is shown above the columns. Bars denote mean ± s.e.m. of three experiments; *P<0.05. Image collected and cropped by CiteAb from the following publication (https://journals.biologists.com/dmm/article/doi/10.1242/dmm.013078/2587…), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Simple Western

20 µg/mL
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

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.

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: 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

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse STAT5a/b Pan Specific Antibody

For research use only

Citations for Human/Mouse STAT5a/b Pan Specific Antibody

Customer Reviews for Human/Mouse STAT5a/b Pan Specific Antibody (1)

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  • Human/Mouse STAT5a/b Pan Specific Antibody
    Name: Anonymous
    Application: Chromatin Immunoprecipitation
    Sample Tested: Alzheimer's disease brain
    Species: Human
    Verified Customer | Posted 06/04/2019

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