Skip to content

Histone H2A.X Recombinant Rabbit Monoclonal Antibody [JM06-42]

Usd: 385

Specification

Bulk Order Quote | Customization Request

Catalog# ET1705-97

Histone H2A.X Recombinant Rabbit Monoclonal Antibody [JM06-42]

Application

  • WB

  • IF-Tissue

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Histone H2A.X Recombinant Rabbit Monoclonal Antibody [JM06-42]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within N-terminal human Histone H2A.X.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Tissue, IHC-P, IP

Molecular Weight

Predicted band size: 15 kDa

Positive Control

Raji cell lysate, MCF-7 cell lysate, human pancreas tissue, mouse testis tissue, rat brain tissue, human stomach carcinoma tissue, mouse pancreas tissue, rat pancreas tissue.

Conjugation

unconjugated

Clone Number

JM06-42

RRID

AB_3070625

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:2000

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:1,000-1:4,000

  • IP

  • Use at an assay dependent concentration.

Target

Function

Histone H2A.X is a member of the Histone H2A family, which is involved in nucleosomal organization of chromatin. The H2AFX gene is located in close proximity to the Porphobilinogen deaminase (PBG-D) gene in both mouse and human, and maps to chromosome 9 and 11q23, respectively. H2A.X meets from the other members of the H2A family by the presence of a highly conserved C-terminal motif. It is wide phosphorylated in response to ionizing radiation and plays an important role in the recognition and repair of DNA fragments are involved in the heavy chain constant region of cells involved in class switch recombination (CSR), a region-specific DNA reaction that replaces one immunoglobulin heavy chain constant region gene with another. The phosphorylated γ-H2A.X is also thought to firing new grades factors, including Rad50, Rad51 and BRCA1.

Background References

1. Bezine E et al. Cell resistance to the Cytolethal Distending Toxin involves an association of DNA repair mechanisms. Sci Rep 6:36022 (2016)

2. Xiong J et al. Stemness factor Sall4 is required for DNA damage response in embryonic stem cells. J Cell Biol 208:513-20 (2015).

Sequence Similarity

Belongs to the histone H2A family.

Post-translational Modification

Phosphorylated on Ser-140 (to form gamma-H2AX or H2AX139ph) in response to DNA double strand breaks (DSBs) generated by exogenous genotoxic agents and by stalled replication forks, and may also occur during meiotic recombination events and immunoglobulin class switching in lymphocytes. Phosphorylation can extend up to several thousand nucleosomes from the actual site of the DSB and may mark the surrounding chromatin for recruitment of proteins required for DNA damage signaling and repair. Widespread phosphorylation may also serve to amplify the damage signal or aid repair of persistent lesions. Phosphorylation of Ser-140 (H2AX139ph) in response to ionizing radiation is mediated by both ATM and PRKDC while defects in DNA replication induce Ser-140 phosphorylation (H2AX139ph) subsequent to activation of ATR and PRKDC. Dephosphorylation of Ser-140 by PP2A is required for DNA DSB repair. In meiosis, Ser-140 phosphorylation (H2AX139ph) may occur at synaptonemal complexes during leptotene as an ATM-dependent response to the formation of programmed DSBs by SPO11. Ser-140 phosphorylation (H2AX139ph) may subsequently occurs at unsynapsed regions of both autosomes and the XY bivalent during zygotene, downstream of ATR and BRCA1 activation. Ser-140 phosphorylation (H2AX139ph) may also be required for transcriptional repression of unsynapsed chromatin and meiotic sex chromosome inactivation (MSCI), whereby the X and Y chromosomes condense in pachytene to form the heterochromatic XY-body. During immunoglobulin class switch recombination in lymphocytes, Ser-140 phosphorylation (H2AX139ph) may occur at sites of DNA-recombination subsequent to activation of the activation-induced cytidine deaminase AICDA. Phosphorylation at Tyr-143 (H2AXY142ph) by BAZ1B/WSTF determines the relative recruitment of either DNA repair or pro-apoptotic factors. Phosphorylation at Tyr-143 (H2AXY142ph) favors the recruitment of APBB1/FE65 and pro-apoptosis factors such as MAPK8/JNK1, triggering apoptosis. In contrast, dephosphorylation of Tyr-143 by EYA proteins (EYA1, EYA2, EYA3 or EYA4) favors the recruitment of MDC1-containing DNA repair complexes to the tail of phosphorylated Ser-140 (H2AX139ph).; Monoubiquitination of Lys-120 (H2AXK119ub) by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression (By similarity). Following DNA double-strand breaks (DSBs), it is ubiquitinated through 'Lys-63' linkage of ubiquitin moieties by the E2 ligase UBE2N and the E3 ligases RNF8 and RNF168, leading to the recruitment of repair proteins to sites of DNA damage. Ubiquitination at Lys-14 and Lys-16 (H2AK13Ub and H2AK15Ub, respectively) in response to DNA damage is initiated by RNF168 that mediates monoubiquitination at these 2 sites, and 'Lys-63'-linked ubiquitin are then conjugated to monoubiquitin; RNF8 is able to extend 'Lys-63'-linked ubiquitin chains in vitro. H2AK119Ub and ionizing radiation-induced 'Lys-63'-linked ubiquitination (H2AK13Ub and H2AK15Ub) are distinct events.; Acetylation at Lys-37 increases in S and G2 phases. This modification has been proposed to play a role in DNA double-strand break repair (By similarity).

Subcellular Location

Nucleus. Chromosome.

UNIPROT

SwissProt: P16104 Human

SwissProt: P27661 Mouse

SwissProt: D3ZXP3 Rat

Synonyms

AW228881 antibody

H2A histone family member X antibody

H2A.FX antibody

H2A.X antibody

H2a/x antibody

H2AFX antibody

H2AX antibody

H2AX histone antibody

H2AX_HUMAN antibody

Hist5.2ax antibody

Expand

Images

  • Western blot analysis of Histone H2A.X on different lysates with Rabbit anti-Histone H2A.X antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/1,000 dilution.<br /><br />Lane 1: Raji cell lysate<br />Lane 2: NIH/3T3 cell lysate<br />Lane 3: C6 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 15 kDa<br />Observed band size: 15 kDa<br /><br />Exposure time: 3 minutes;<br /><br />15% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Histone H2A.X on different lysates with Rabbit anti-Histone H2A.X antibody (ET1705-97) at 1/1,000 dilution.

    Lane 1: Raji cell lysate
    Lane 2: NIH/3T3 cell lysate
    Lane 3: C6 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 15 kDa
    Observed band size: 15 kDa

    Exposure time: 3 minutes;

    15% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-97) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunofluorescence staining of paraffin- embedded human pancreas tissue using anti-Histone H2A.X antibody.The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with <a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a> at 1/50 dilution for 10 hours at 4℃ and detected using Alexa Fluor&reg; 488 conjugate-Goat anti-Rabbit IgG (H+L) Secondary Antibody at a dilution of 1:500 for 1 hour at room temperature.

    Immunofluorescence staining of paraffin- embedded human pancreas tissue using anti-Histone H2A.X antibody.The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with ET1705-97 at 1/50 dilution for 10 hours at 4℃ and detected using Alexa Fluor® 488 conjugate-Goat anti-Rabbit IgG (H+L) Secondary Antibody at a dilution of 1:500 for 1 hour at room temperature.

  • Immunofluorescence staining of paraffin- embedded mouse testis tissue using anti-Histone H2A.X antibody.The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with <a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a> at 1/50 dilution for 10 hours at 4℃ and detected using Alexa Fluor&reg; 488 conjugate-Goat anti-Rabbit IgG (H+L) Secondary Antibody at a dilution of 1:500 for 1 hour at room temperature.

    Immunofluorescence staining of paraffin- embedded mouse testis tissue using anti-Histone H2A.X antibody.The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with ET1705-97 at 1/50 dilution for 10 hours at 4℃ and detected using Alexa Fluor® 488 conjugate-Goat anti-Rabbit IgG (H+L) Secondary Antibody at a dilution of 1:500 for 1 hour at room temperature.

  • Immunofluorescence staining of paraffin- embedded rat brain tissue using anti-Histone H2A.X antibody.The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with <a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a> at 1/50 dilution for 10 hours at 4℃ and detected using Alexa Fluor&reg; 488 conjugate-Goat anti-Rabbit IgG (H+L) Secondary Antibody at a dilution of 1:500 for 1 hour at room temperature.

    Immunofluorescence staining of paraffin- embedded rat brain tissue using anti-Histone H2A.X antibody.The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with ET1705-97 at 1/50 dilution for 10 hours at 4℃ and detected using Alexa Fluor® 488 conjugate-Goat anti-Rabbit IgG (H+L) Secondary Antibody at a dilution of 1:500 for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue with Rabbit anti-Histone H2A.X antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human pancreas tissue with Rabbit anti-Histone H2A.X antibody (ET1705-97) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-97) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Histone H2A.X antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Histone H2A.X antibody (ET1705-97) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-97) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Histone H2A.X antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Histone H2A.X antibody (ET1705-97) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-97) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue with Rabbit anti-Histone H2A.X antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue with Rabbit anti-Histone H2A.X antibody (ET1705-97) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-97) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue with Rabbit anti-Histone H2A.X antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/4,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/4,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue with Rabbit anti-Histone H2A.X antibody (ET1705-97) at 1/4,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-97) at 1/4,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat pancreas tissue with Rabbit anti-Histone H2A.X antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/4,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-97" style="font-weight: bold;text-decoration: underline;">ET1705-97</a>) at 1/4,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat pancreas tissue with Rabbit anti-Histone H2A.X antibody (ET1705-97) at 1/4,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-97) at 1/4,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation