Alexa Fluor® 647 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
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Product Citations
publications
W16171A_A647_H2AX_Antibody_1_110119
HeLa cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with Triton X-100 for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 1 µg/mL (1:500 dilution) of either Alexa Fluor® 647 Rat IgG2a, κ Isotype Ctrl Antibody (Cat. No. 400526, panel A) or Alexa Fluor® 647 anti-H2A.X antibody (panel B) overnight at 4°C. Nuclei were counterstained with DAPI and the image was captured with a 60X objective.
  • W16171A_A647_H2AX_Antibody_1_110119
    HeLa cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with Triton X-100 for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 1 µg/mL (1:500 dilution) of either Alexa Fluor® 647 Rat IgG2a, κ Isotype Ctrl Antibody (Cat. No. 400526, panel A) or Alexa Fluor® 647 anti-H2A.X antibody (panel B) overnight at 4°C. Nuclei were counterstained with DAPI and the image was captured with a 60X objective.
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600205 25 µg £109
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600206 100 µg £254
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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 and conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

ICC - Quality tested

Recommended Usage

Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633 nm / 635 nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 nm)
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_2820097 (BioLegend Cat. No. 600205)
AB_2820097 (BioLegend Cat. No. 600206)

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: 1    Revision Date: 11/01/2019

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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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.
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