AF2697

antibody from R&D Systems

Targeting: NFKB1 KBF1, NF-kappaB, NF-kB1, NFkappaB, NFKB-p50, p105, p50

Western blot (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

AF2697

Provider

R&D Systems

Product name

Human/Mouse NFkB1 Antibody

Antibody type

Polyclonal

Description

Antigen Affinity-purified. Detects endogenous human and mouse NF kappa B1 in Western blots.

Reactivity

Human, Mouse

Host

Goat

Conjugate

Unconjugated

Antigen sequence

NP_003989


Isotype

IgG

Vial size

100 ug

Concentration

LYOPH

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.

Western blot

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Human NF kappa B1 by Western Blot. Western blot shows lysates of Raji human Burkitt's lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse NF kappa B1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2697) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for NF kappa B1 at approximately 50 and 105 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of NF kappa B1-regulated Genes by Chromatin Immunoprecipitation. Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin overnight was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. NF kappa B1/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human/Mouse NF kappa B1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2697) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The bcl-2 promoter was detected by standard PCR. This application has not been tested in mouse samples.