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Purified anti-H2A.X Antibody

Pricing & Availability
Clone
W16171A (See other available formats)
Regulatory Status
RUO
Other Names
H2A.X Variant Histone, H2A Histone Family Member X, Histone H2A.X, Histone H2AX, H2AFX
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
1. W16171A_PURE_H2AX_Antibody_1_092917
Total cell lysates (15 µg protein) from HeLa (lane 1), HEK293T (lane 2), MCF-7 (lane 3), and NIH/3T3 (lane 4) cells were resolved by 4-20% Tris-Glycine gel electrophoresis, transferred to nitrocellulose, and probed with 1 µg/mL (1:500 dilution) of purified anti-H2A.X antibody (Clone W16171A, left) or a competitor’s clone used at the manufacturer’s recommended dilution. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG (cat #405405) for clone W161171A and HRP donkey anti-rabbit IgG (cat #406401) for the competitor’s clone, each at a 1:3000 dilution. Membranes were then probed with purified anti-GAPDH antibody (lower, cat# 631402) to confirm equal protein loading. Lane M: MW ladder.
  • 1. W16171A_PURE_H2AX_Antibody_1_092917
    Total cell lysates (15 µg protein) from HeLa (lane 1), HEK293T (lane 2), MCF-7 (lane 3), and NIH/3T3 (lane 4) cells were resolved by 4-20% Tris-Glycine gel electrophoresis, transferred to nitrocellulose, and probed with 1 µg/mL (1:500 dilution) of purified anti-H2A.X antibody (Clone W16171A, left) or a competitor’s clone used at the manufacturer’s recommended dilution. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG (cat #405405) for clone W161171A and HRP donkey anti-rabbit IgG (cat #406401) for the competitor’s clone, each at a 1:3000 dilution. Membranes were then probed with purified anti-GAPDH antibody (lower, cat# 631402) to confirm equal protein loading. Lane M: MW ladder.
  • 2. W16171A_PURE_H2AX_Antibody_2_092917
    HeLa cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 1:100 (5 µg/mL), 1:200 (2.0 µg/mL) and 1:500 (1.0 µg/mL) dilutions of either rat IgG or purified anti-H2A.X antibody for two hours at room temperature, followed by incubation with Alexa Fluor® 594 goat anti-rat IgG (cat #405422) at 2.0 µg/mL. Nuclei were counterstained with DAPI, and the image was captured with a 60X objective.
  • 3. W16171A_PURE_H2AX_Antibody_061022
    IHC staining using purified anti-H2A.X antibody (clone W16171A) on formalin-fixed paraffin-embedded human cerebellum tissue. The tissue was incubated with 10 μg/mL of anti-H2A.X antibody overnight at 4°C, followed by incubation with 2.5 μg/mL of Alexa Fluor® 647 goat anti-rat IgG antibody (red) (Cat. No. 405416) for one hour at room temperature. Slide was mounted with ProLong™ Gold Antifade Mountant. Nuclei were counterstained with DAPI (blue, right panel) (Cat. No. 422801). The image was captured with a 40x objective. Scalebar = 50 μM
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600201 25 µg £70
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600202 100 µg £174
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Description

Histone subunit H2A, along with subunits 2B, 3, and 4, make up the core subunits of the nucleosome octomer. An octomer contains two protomers of each subunit tightly wrapped around a ~147 bp segment of DNA. Histones have integral roles in chromatin integrity, genomic stability, and gene regulation. Post-translational modification of histones in response to certain stimuli results in alterations of nucleosomal positioning relative to DNA. Histone H2A.X is a non-allelic variant of Histone 2A that harbors a C-terminal extension and is essential for checkpoint mediated cell cycle arrest and DNA double-stranded break (DSB) repair in response to both endogenous and exogenous agents, as well as meiotic recombination events and immunoglobulin class switching in lymphocytes. Phosphorylation of C-terminal residue serine 139 by ATM (γ-H2A.X) results in the recruitment of DSB-repair machinery. Phopshorylation of H2A.X is also critical for chromatin fragmentation during apoptosis.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Synthetic human histone H2A.X peptide (127-143) conjugated to KLH.
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, IHC-P - Verified
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 1.0 - 5.0 µg per ml (1:100-1:500 dilution). For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/ml (1:100-1:500 dilution) is recommended. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 2.0 - 10.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This product is a monoclonal antibody raised against the C-terminus of H2A.X (residues 127-143); BioLegend’s existing antibody against H2A.X (Poly6133, cat# 613302) is a polyclonal antibody which was generated against (partial), N-terminal H2A.X.

This clone is not recommended for ChIP (Chromatin Immunoprecipitation) assays (as determined by in-house testing).

RRID
AB_2716198 (BioLegend Cat. No. 600201)
AB_2716199 (BioLegend Cat. No. 600202)

Antigen Details

Structure
Histone H2A.X is a 143 amino acid protein with a predicted molecular weight of 15.1 kD.
Distribution

Ubiquitous tissue expression; nuclear localization

Function
H2A.X, upon phosphorylation, promotes DNA repair and maintains genomic stability. Important for recombination between immunoglobulin switch regions.
Interaction
ATM, MDC1, TP53BP1, BRCA1, MRE11, RAD50, NBN
Biology Area
Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Cell Cycle/DNA Replication, Chromatin Remodeling/Epigenetics, DNA Repair/Replication
Antigen References
  1. Chen CC, et al. 2017. Proc. Natl. Acad. Sci. 114: 7665.
  2. Natale F, et al. 2017. Nat. Commun. 8: 15760.
  3. Bhargava R, et al. 2017. Proc. Natl. Acad. Sci. 114: 728.
  4. Weyemi U, et al. 2016. Nat. Commun. 7: 10711.
  5. Rezaeian AH, et al. 2017. Nat. Cell. Biol. 19: 38.
  6. Horn S, et al. 2015. Biochim. Biophys. Acta. 1853: 2199.
  7. Reina-San-Martin B, et al. 2003. J. Exp. Med. 197:1767
  8. Celeste A, et al. 2002. Science 296:922.
  9. Mannironi C, et al.1989. Nucleic Acids Res. 17:9113.
Gene ID
3014 View all products for this Gene ID
UniProt
View information about H2A.X on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 2    Revision Date: 06/10/2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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