MA5-24696

antibody from Invitrogen Antibodies

Targeting: MACROH2A1 H2AFY, macroH2A1.2

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-24696

Provider

Invitrogen Antibodies

Product name

Histone Macro-H2A.1 Recombinant Rabbit Monoclonal Antibody (RM248)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody reacts to the Core histone macro-H2A.1 (Histone macroH2A1) protein, independent of post-translational modifications. No cross reactivity with other histone proteins. Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Antibody clone number

RM248

Vial size

100 µg

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on acid extracts (20 µg lysate) of HeLa (Lane 1), K-562 (Lane 2), U-2 OS (Lane 3), Hep G2 (Lane 4) and NIH/3T3 (Lane 5). The blot was probed with Anti-Core Histone Macro-H2A.1 Monoclonal Antibody (Product # MA5-24696, 1 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 39 kDa band corresponding to Core Histone Macro-H2A.1 was observed across the cell lines tested.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Core Histone Macro-H2A.1 was performed using 70% confluent log phase HeLa cells. 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 Core Histone Macro-H2A.1 (Product # MA5-24696) 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 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 Methyl-Histone H3 (Lys9) protein at specific gene loci using Anti-Methyl-Histone H3 (Lys9) Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-Methyl-Histone H3 (Lys9) Polyclonal Antibody (Product # PA5-11183, 3 µg) on sheared chromatin from 2 million 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 MYOD gene used as positive and c-Fos, GAPDH, SAT2 satellite repeats used as negative target genes/binding sites. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.