701108

antibody from Invitrogen Antibodies

Targeting: STAT5A MGF, STAT5

Western blot (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

701108

Provider

Invitrogen Antibodies

Product name

STAT5 alpha Recombinant Rabbit Monoclonal Antibody (6H2L9)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

6H2L9

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of STAT5 alpha in whole cell extracts of serum-starved HeLa cells treated with IFN-a (150 ng/mL, 15 min) using a STAT5 alpha recombinant rabbit monoclonal antibody (Product # 701108) at a dilution of 2.5 µg/mL. Samples were detected using chemiluminescence (ECL). Results show a band at ~90 kDa.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of STAT5A was done on Jurkat cells treated with IFN gamma (150 ng/ml, 15 minutes). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Tritonª X-100 for 20 minutes, and blocked with 5% BSA for 1 hour at room temperature. Cells were labeled with ABfinityª STAT5A Recombinant Rabbit Monoclonal Antibody (701108, red histogram) or with rabbit isotype control (pink histogram) at 2 µg-4 µg/million cells in 2.5% BSA. After incubation at room temperature for 2-3 hours, the cells were labeled with Alexa Fluor¨ 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune¨ Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

ChIP- qPCR analysis of STAT5A was performed with 3 µg/mL of the STAT5A Recombinant Rabbit Monoclonal Antibody (Product # 701108) on sheared chromatin from 2 million HeLa cells treated with IFN-gamma (50 ng/mL) for 1h using the MAGnify Chromatin Immunoprecipitation System (Product # 49-2024). Normal rabbit IgG (3 µg/mL) was used as a negative IP control. The purified DNA from each ChIP sample was analyzed by StepOnePlus Real-Time PCR System (Product # 4376600) with primers for the promoter of active AP-1 and COX2 gene, used as positive control targets, and the GAPDH gene, used as negative control target. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.