Invitrogen

Phospho-Histone H2A.X (Ser139) Monoclonal Antibody (CR55T33), eBioscience™

Phospho-Histone H2A.X (Ser139) Antibody in Immunocytochemistry (ICC/IF)

图: 1 / 11

Phospho-Histone H2A.X (Ser139) Antibody (14-9865-82) in ICC/IF

Immunofluorescence analysis of phospho-H2A.X was performed using 70% confluent log phase HeLa cells treated with hydroxyurea (3 mM for 6 hr). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Phospho-Histone H2A.X (Ser139) Monoclonal Antibody (CR55T33), eB... View More

Phospho-Histone H2A.X (Ser139) Antibody in Immunohistochemistry (Paraffin) (IHC (P))

Phospho-Histone H2A.X (Ser139) Antibody in Western Blot (WB)

Phospho-Histone H2A.X (Ser139) Antibody in Flow Cytometry (Flow)

Phospho-Histone H2A.X (Ser139) Antibody in Immunohistochemistry (IHC)

Product Image Gallery

产品信息

14-9865-82

应用

建议稀释比

已发表文章

免疫印迹 (WB)

1:500

查看2篇已发表文章 2篇已发表文章

免疫组化 (IHC)

查看1篇已发表文章 1篇已发表文章

免疫组化（石蜡） (IHC (P))

5 µg/mL

查看1篇已发表文章 1篇已发表文章

免疫细胞化学 (ICC/IF)

5 µg/mL

查看1篇已发表文章 1篇已发表文章

流式细胞分析 (Flow)

0.03 µg/test

查看2篇已发表文章 2篇已发表文章

产品规格

种属反应

Human, Mouse

已发表种属

Human

宿主/亚型

Mouse / IgG1, kappa

分类

Monoclonal

类型

Antibody

克隆号

CR55T33

偶联物

Unconjugated Unconjugated Unconjugated

形式

Liquid

浓度

0.5 mg/mL

纯化类型

Affinity chromatography

保存液

PBS, pH 7.2

内含物

0.09% sodium azide

保存条件

4° C

运输条件

Ambient (domestic); Wet ice (international)

AB_2573048

产品详细信息

Description: The CR55T33 monoclonal antibody recognizes phosphorylated serine 139 of human and mouse H2AX. H2AX is a member of the H2A histone family that complex with DNA and other histones to form the repeating nucleosome units characteristic of eukaryotic chromatin. Nucleosomes consist of approximately 147 base pairs of DNA wrapped around an octamer of histones composed of two each of the four histone proteins: H2A, H2B, H3 and H4. After induction of DNA damage such as double-strand breaks by irradiation, genotoxic stresses, replication errors or gene recombination, PI3K-like kinases (e.g., ataxia telangiectasia mutated (ATM), ataxia telangiectasia Rad-3-related (ATR), and DNA-dependent protein kinase (DNA-PK) are activated to phosphorylate serine 139 in H2AX. This early phosphorylation event plays a critical role in recruiting proteins involved in DNA repair.

The monoclonal antibody CR55T33 recognizes a single band of approximately 15 kDa on reduced cell lysates from Jurkat cells stimulated with etoposide.

Applications Reported: This CR55T33 antibody has been reported for use in intracellular staining followed by flow cytometric analysis, western blotting, immunohistochemical staining of formalin-fixed paraffin embedded tissue sections, and immunocytochemistry (fluorochrome-conjugated CR55T33 is recommended for use in intracellular flow cytometry).

Applications Tested: The CR55T33 has been tested by immunocytochemistry of methanol-fixed cells and by immunohistochemistry of human tissue using either low or high pH antigen retrieval and can be used at 5 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. This CR55T33 antibody has also been tested by intracellular staining and flow cytometric analysis of stimulated Jurkat cells using the Foxp3/Transcirption Factor Buffer Set (Product # 00-5523-00) and protocol. This can be used at 0.03 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Protocols: We recommend Protocol B: One-step protocol: intracellular (nuclear) proteins. Alternatively, Protocol C: Two-step protocol: Fixation/Methanol can also be used. Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins cannot be used. All Protocols can be found in the "Staining intracellular Antigens for Flow Cytometry Protocol" located in the Best Protocols Section under the Resources tab online.

eFluor® 660 is a replacement for Alexa Fluor® 647. eFluor® 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome.

Purity: Greater than 90%, as determined by SDS-PAGE.

Aggregation: Less than 10%, as determined by HPLC.

Filtration: 0.2 µm post-manufacturing filtered.

靶标信息

Histone H2A.X (H2AX) is a member of the histone H2A family which is one of the four core histones making up the nucleosome core particle. In eukaryotes, DNA double strand breaks (DSBs) have been shown to trigger the phosphorylation of serine 139 at the carboxy terminus of histone H2AX resulting in gamma-H2AX. The phosphorylation of H2AX can be detected by Western blotting or immunofluorescence, revealing the frequency of DSBs. The phosphatidylinositol 3-kinases have been implicated in H2AX phosphorylation, but it is unclear if ATM is the primary H2AX kinase or if other members of the family such as DNA-PK and ATR contribute in a similar manner. Structurally, H2A.x contains 143 amino acid residues. Histone H2A.X is considered a basal histone, being synthesized in G1 as well as in S-phase, and its mRNA contains polyA addition motifs and a polyA tail along with the conserved stem-loop and U7 binding sequences involved in the processing and stability of replication type histone mRNAs. There are two forms of Histone H2A.X mRNA, one about 1600 bases long and contains polyA; the other about 575 bases long, lacking polyA. The short form behaves as a replication type histone mRNA, while the longer behaves as a basal type histone mRNA. Histone H2A.X maps to the 11q23.2-q23.3 region of the human chromosome. Histone H2A.x contributes to histone-formation and therefore the structure of DNA. Histone H2A variant H2A.x specifically regulates the interaction of MDC1 (mediator of DNA damage checkpoint protein 1), a DNA repair protein to the sites of DNA damage.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

Product References

The mitotic checkpoint is a targetable vulnerability of carboplatin-resistant triple negative breast cancers.

Scientific reports

Moens S,Zhao P,Baietti MF,Marinelli O,Van Haver D,Impens F,Floris G,Marangoni E,Neven P,Annibali D,Sablina AA,Amant F

Published figure using Phospho-Histone H2A.X (Ser139) monoclonal antibody (Product # 14-9865-82) in Flow Cytometry

Thu Feb 04 00:00:00 UTC 2021

Synergistic lethality between PARP-trapping and alantolactone-induced oxidative DNA damage in homologous recombination-proficient cancer cells.

Oncogene

Wang H,Zhang S,Song L,Qu M,Zou Z

Published figure using Phospho-Histone H2A.X (Ser139) monoclonal antibody (Product # 14-9865-82) in Western Blot

Wed Apr 01 00:00:00 UTC 2020

Synergistic lethality between PARP-trapping and alantolactone-induced oxidative DNA damage in homologous recombination-proficient cancer cells.

Oncogene

Wang H,Zhang S,Song L,Qu M,Zou Z

Published figure using Phospho-Histone H2A.X (Ser139) monoclonal antibody (Product # 14-9865-82) in Western Blot

Wed Apr 01 00:00:00 UTC 2020

Evaluation of cisplatin-induced injury in human kidney organoids.

American journal of physiology. Renal physiology

Digby JLM,Vanichapol T,Przepiorski A,Davidson AJ,Sander V

14-9865-80 was used in Immunohistochemistry (Paraffin) to characterize induced pluripotent stem cell-derived human kidney organoids and their response to cisplatin, a chemotherapeutic drug that induces AKI and preferentially damages the proximal tubule.

Wed Apr 01 00:00:00 UTC 2020

MAU2 and NIPBL Variants Impair the Heterodimerization of the Cohesin Loader Subunits and Cause Cornelia de Lange Syndrome.

Cell reports

Parenti I,Diab F,Gil SR,Mulugeta E,Casa V,Berutti R,Brouwer RWW,Dupé V,Eckhold J,Graf E,Puisac B,Ramos F,Schwarzmayr T,Gines MM,van Staveren T,van IJcken WFJ,Strom TM,Pié J,Watrin E,Kaiser FJ,Wendt KS

14-9865-80 was used in Immunocytochemistry to highlight a novel mechanism protective against out-of-frame mutations that is potentially relevant for other genetic conditions.