Phospho-Histone H2A.X (Ser139) Recombinant antibody

Phospho-Histone H2A.X (Ser139) Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IHC, IF/ICC, ELISA

Conjugate

Unconjugated

Publications

1

CloneNo.

5N19

Cat no : 83307-2-RR

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 83307-2-RR

  Non-treated Jurkat cells, and staurosporine treated Jurkat cells were subjected to SDS PAGE followed by western blot with 83307-2-RR (Phospho-Histone H2A.X (Ser139) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Alpha Tubulin (66031-1-Ig) antibody as loading control.

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  IHC staining of Jurkat using 83307-2-RR

  Immunohistochemical analysis of paraffin-embedded Jurkat cells slide using 83307-2-RR (Phospho-Histone H2A.X (Ser139) antibody) at dilution of 1:4000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 83307-2-RR

  Immunofluorescent analysis of (4% PFA) fixed UV treated HeLa cells using Phospho-Histone H2A.X (Ser139) antibody (83307-2-RR, Clone: 5N19 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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Tested Applications

• Positive WB detected in: Staurosporine treated Jurkat cells
• Positive IHC detected in: Jurkat cells; 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: UV treated HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:2000-1:8000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800

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

83307-2-RR targets Phospho-Histone H2A.X (Ser139) in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: Peptide
• Full Name: H2A histone family, member X
• Calculated Molecular Weight: 15 kDa
• Observed Molecular Weight: 15 kDa
• GenBank Accession Number: BC013416
• Gene Symbol: Histone H2A.X
• Gene ID (NCBI): 3014
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• 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

The histone variant H2AX is a major component of the DNA damage response (DDR), especially functioning in amplifying DNA damage signals. In response to DNA double-strand breaks (DSBs), H2AX is instantaneously phosphorylated at Ser139 (a form called cH2AX) by the kinases ATM and ATR. The phosphorylation of H2AX at Ser139, resulting in the formation of gamma-H2AX puncta in the nuclei, is an early event in the cellular response to DNA damage. Therefore, phospho-Histone H2A. X (Ser139) is also known as γH2AX. The phosphorylation site of H2AX, Ser139, has also been described as Ser140 in other literature, and they recognize the same amino acid site. (PMID: 22908299, PMID: 30106130, PMID:22941631)

Publications

Species	Application	Title
human	WB	Transl Oncol; PiR-hsa-23533 promotes malignancy in head and neck squamous cell carcinoma via USP7; Authors - Hanlin Hu