Histone H2A.X Monoclonal antibody

Histone H2A.X Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

1

CloneNo.

3E11B2

Cat no : 68888-1-Ig

Synonyms

H2AFX, Histone H2A, H2AX, H2A.X, H2A histone family, member X

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Validation Data Gallery

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  WB analysis using 68888-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 68888-1-Ig (Histone H2A.X antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  IHC staining of human stomach cancer using 68888-1-Ig

  Immunohistochemical analysis of paraffin-embedded human stomach cancer slide using 68888-1-Ig (Histone H2A.X antibody) at dilution of 1:1000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 68888-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using Histone H2A.X antibody (68888-1-Ig, Clone: 3E11B2 ) at dilution of 1:1000 and Multi-rAb CoraLite® Plus 594-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (Cat.NO. RGAM004 ).

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  IF Staining of HepG2 using 68888-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using Histone H2A.X antibody (68888-1-Ig, Clone: 3E11B2 ) at dilution of 1:1000 and Multi-rAb CoraLite® Plus 594-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (Cat.NO. RGAM004 ).

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  FC experiment of HepG2 using 68888-1-Ig

  1x10^6 HepG2 cells were intracellularly stained with 0.1 ug Histone H2A.X Monoclonal antibody (68888-1-Ig, Clone:3E11B2) and Multi-rAb CoraLite ® Plus 647-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (RGAM005)(red), or 0.1 ug Mouse IgG1 isotype control Mouse McAb (66360-1-Ig, Clone: 1F8D3) (blue). Cells were fixed with 4% PFA and permeabilized with 0.1% TritonX-100.

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Tested Applications

• Positive WB detected in: Saos-2 cells, MDA-MB-231 cells, HeLa cells, LNCaP cells, HEK-293 cells, Jurkat cells, HSC-T6 cells, NIH/3T3 cells, 4T1 cells
• Positive IHC detected in: human stomach cancer; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:500-1:2000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.10 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 1 publications below

Product Information

68888-1-Ig targets Histone H2A.X in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: H2A histone family, member X
• Calculated Molecular Weight: 15 kDa
• Observed Molecular Weight: 17 kDa
• GenBank Accession Number: BC013416
• Gene Symbol: Histone H2A.X
• Gene ID (NCBI): 3014
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Histone H2A.X belongs to the histone H2A family, which is synthesized in G1 and S phases. It is involved in nucleosomal organization of chromatin together with other histone proteins and is especially important for recombination between immunoglobulin switch regions. H2A.X becomes phosphorylated on serine 139 (to form gamma-H2AFX or H2AX139ph) in response to DNA double strand breaks (DSBs) generated by exogenous genotoxic agents and by stalled replication forks, which promotes DNA repair and maintains genomic stability. The calculated molecular weight of H2AX is 15 kDa, but the ubiquitinated H2A.X is about 22 kDa.

Publications

Species	Application	Title
human	WB	J Med Chem; Discovery of Indolo[3,2-c]isoquinoline Derivatives as Novel Top1/2 Dual Inhibitors with Orally Efficacious Antitumor Activity and Low Toxicity; Authors - Bichuan Wang