MA5-24665

antibody from Invitrogen Antibodies

Targeting: H2AZ2 H2AFV, H2AV, MGC10170, MGC10831, MGC1947

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-24665

Provider

Invitrogen Antibodies

Product name

H2A.ZK7ac Recombinant Rabbit Monoclonal Antibody (RM222)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody reacts to Histone H2A.Z acetylated at Lysine 7 (K7ac). No cross reactivity with non-modified Lysine 7 or other acetylated Lysines in histone H2A.

Antibody clone number

RM222

Concentration

1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of H2A.Z and H2A.ZK7AC was performed by loading 0.5 ng of acid purified histone protein and PageRuler Plus Protein Ladder (Product # 26619) onto a 30% Acrylamide/Bis gel. Proteins were transferred and membrane was blocked in 5% BSA for one hour at room temperature. H2A.Z and H2A.ZK7AC was detected at approximately 14 kDa using a H2A.Z monoclonal antibody (Product # MA5-24666) or H2A.ZK7AC (Product # MA5-24665) at a dilution of 1:5,000 in 5% BSA overnight at 4°C on a rocking platform, followed by a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5,000 for one hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076) and the myECL Imager (Product # 62236). Data Courtesy of Wei Xu’s laboratory at the University of Wisconsin Madison.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot of acid extracts from HeLa cells treated (+) or untreated (-) with sodium butyrate, using Acetyl-Histone H2A.Z (Lys7) Antibody (Product # MA5-24665) at 0.5 mg/mL.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of HeLa cells treated with sodium butyrate, using Acetyl-Histone H2A.Z (Lys7) Antibody (Product # MA5-24665) (red). Actin filaments have been labeled with FITC-conjugated phalloidin (green).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Acetyl-Histone H2A.Z (Lys7) was performed using 70% confluent log phase HeLa cells treated with sodium butyrate. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Acetyl-Histone H2A.Z (Lys7) Rabbit Monoclonal Antibody (RM222) (Product # MA5-24665) at 5 microgram/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents untreated cells with relatively lower expression of Acetyl-Histone H2A.Z (Lys7) (RM222). Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Enrichment of endogenous Acetyl-Histone H2A.Z (Lys7) protein at specific gene loci using Anti-Acetyl-Histone H2A.Z (Lys7) Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-Acetyl-Histone H2A.Z (Lys7) Mouse Monoclonal Antibody (Product # MA5-24665, 3 µg) on sheared chromatin from 2 million Sodium butyrate-treated HeLa cells using the "MAGnify ChIP system" kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over the ACTB gene (active) and SAT2 satellite repeats (inactive). A schematic diagram of the ACTB gene is shown on top of the figure. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Acetyl-Histone H2A.Z (Lys7) Antibody (Product # MA5-24665) specifically reacts to Histone H2A.Z acetylated at Lysine 7 (K7ac). No cross reactivity with non-modified Lysine 7 or other acetylated Lysines in histone H2A.