Invitrogen

Phospho-Histone H2A.X (Ser139) Monoclonal Antibody (CR55T33), eFluor™ 660, eBioscience™

Name: Phospho-Histone H2A.X (Ser139) Monoclonal Antibody (CR55T33), eFluor\u2122 660, eBioscience\u2122
Brand: Invitrogen
SKU: 50-9865-42

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6 篇参考文献

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说明书

实验方案

问题与答案

技术支持

说明书

实验方案

问题与答案

技术支持

说明书

实验方案

问题与答案

技术支持

抗体检测数据 (3)
已发表图片 (5)

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

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Phospho-Histone H2A.X (Ser139) Antibody (50-9865-42) 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), eF... View More

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)

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产品信息

50-9865-42

应用

建议稀释比

已发表文章

免疫印迹 (WB)

1:400

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免疫组化 (IHC)

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免疫细胞化学 (ICC/IF)

1:50

流式细胞分析 (Flow)

5 µL (0.125 µg)/test

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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

eFluor™ 660 eFluor™ 660 eFluor™ 660

激发/发射光谱

651/668 nm 查看光谱

形式

Liquid

浓度

5 µL/Test

纯化类型

Affinity chromatography

保存液

PBS, pH 7.2, with BSA

内含物

0.09% sodium azide

保存条件

4°C, store in dark, DO NOT FREEZE!

运输条件

Ambient (domestic); Wet ice (international)

AB_2574399

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, and immunocytochemistry.

Applications Tested: This CR55T33 antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of treated human peripheral blood cells using the Foxp3/Transcription Factor Buffer Set (Product # 00-5523-00) and protocol. This can be used at 5 µL (0.125 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. The CR55T33 antibody has also been tested by immunocytochemistry of methanol-fixed human cells and can be used at less than or equal to 10 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

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 BestProtocols® 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 fluorochrome.

Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser.

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.

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