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50ul/100ul/200ul
| 规格: | 50ul | 产品价格: | ¥1180.0 |
|---|---|---|---|
| 规格: | 100ul | 产品价格: | ¥1980.0 |
| 规格: | 200ul | 产品价格: | ¥2800.0 |
| 产品编号 | bs-3185R |
| 英文名称 | phospho-H2AX (Ser139) Rabbit pAb |
| 中文名称 | 磷酸化组蛋白H2AX抗体 |
| 英文别名 | phospho-Histone H2A.X(Ser140); H2AX_HUMAN; 2A.X variant histone; H2A.X; H2A/X; H2AFX; Histone H2AX; |
| 产品应用 | WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=2ug/Test Not yet tested in other applications. |
| 交叉反应 | Human, Mouse, Rat (Dog, Pig, Cow, Horse, Rabbit) |
| 抗体来源 | Rabbit |
| 免疫原 | KLH conjugated Synthesised phosphopeptide derived from human Histone H2AX around the phosphorylation site of Tyr139 |
| 亚型 | IgG |
| 性状 | Liquid |
| 纯化方法 | affinity purified by Protein A |
| 克隆类型 | Polyclonal |
| 理论分子量 | 16 kDa |
| 浓度 | 1mg/ml |
| 储存液 | 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. |
| 研究领域 | Epigenetics and Nuclear Signaling > ChIP assays > ChIP antibodies Epigenetics and Nuclear Signaling > DNA / RNA > DNA Damage & Repair > DNA Damage Response > DNA Damage Recognition Epigenetics and Nuclear Signaling > Histones > H2A > Unmodified Epigenetics and Nuclear Signaling > Histones > Variants |
| 亚基 | The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. Interacts with numerous proteins required for DNA damage signaling and repair when phosphorylated on Ser-140. These include MDC1, TP53BP1, BRCA1 and the MRN complex, composed of MRE11A, RAD50, and NBN. Interaction with the MRN complex is mediated at least in part by NBN. Also interacts with DHX9/NDHII when phosphorylated on Ser-140 and MCPH1 when phosphorylated at Ser-140 or Tyr-143. Interacts with ARRB2; the interaction is detected in the nucleus upon OR1D2 stimulation. |
| 亚细胞定位 | Nucleus. Chromosome. |
| 翻译后修饰 | Phosphorylated on Ser-140 (to form gamma-H2AFX 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)._x000D_
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._x000D_ 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. |
| 相似性 | Belongs to the histone H2A family. |
| 功能 | Variant histone H2A which replaces conventional H2A in a subset of nucleosomes. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Required for checkpoint-mediated arrest of cell cycle progression in response to low doses of ionizing radiation and for efficient repair of DNA double strand breaks (DSBs) specifically when modified by C-terminal phosphorylation. |
| 保存条件 | Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. |
| 注意事项 | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. |
| 背景资料 | Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene encodes a member of the histone H2A family, and generates two transcripts through the use of the conserved stem-loop termination motif, and the polyA addition motif. [provided by RefSeq, Jul 2008]. |
| 应用 | 推荐稀释比例 |
| {WB} | {1:500-2000} |
| {IHC-P} | {1:100-500} |
| {IHC-F} | {1:100-500} |
| {IF} | {1:100-500} |
| {Flow-Cyt} | {2ug/Test} |

_x000D_ Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
_x000D_ Incubation: Anti-Phospho-Histone H2A.X(Ser139) Polyclonal Antibody, Unconjugated(bs-3185R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
_x000D_

_x000D_ Primary Antibody (green line): Rabbit Anti-Phospho-Histone H2A.X (Ser139) antibody (bs-3152R)
_x000D_ Dilution: 1μg /10^6 cells;
_x000D_ Isotype Control Antibody (orange line): Rabbit IgG .
_x000D_ Secondary Antibody : Goat anti-rabbit IgG-APC
_x000D_ Dilution: 1μg /test.
_x000D_ Protocol
_x000D_ The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed._x000D_

_x000D_ Primary Antibody (green line): Rabbit Anti-Phospho-Histone H2A.X (Ser139) antibody (bs-3185R)
_x000D_ Dilution: 2μg /10^6 cells;
_x000D_ Isotype Control Antibody (orange line): Rabbit IgG .
_x000D_ Secondary Antibody : Goat anti-rabbit IgG-FITC
_x000D_ Dilution: 1μg /test.
_x000D_ Protocol
_x000D_ The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed._x000D_

_x000D_ Lane 1: Mouse BV2 cell lysates
_x000D_ Lane 2: Rat Testis tissue lysates
_x000D_ Lane 3: Human 293T cell lysates
_x000D_ Lane 4: Human Jurkat cell lysates
_x000D_ Primary: Anti-Phospho-H2AX (Ser139) (bs-3185R) at 1/5000 dilution
_x000D_ Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
_x000D_ Predicted band size: 16 kDa
_x000D_ Observed band size: 15 kDa
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文献和实验INTRODUCTION In cells and tissues, the histone proteins that constitute the nucleosomes can present multiple post-translational modifications, such as lysine acetylation, lysine and arginine methylation, serine phosphorylation
Gregory, R.I., Randall, T.E., Johnson, C.A., Khosla, S., Hatada, I., O'Neill, L.P., Turner, B.M., and Feil, R. 2001. DNA methylation is linked to deacetylation of histone H3, but not H4, on the imprinted genes Snrpn and U2af1-rs1. Mol. Cell. Biol. 21: 5426
TBS 和 5% NGS 中封闭样品。市售的含有酪蛋白的封闭溶液与磷酸化的一抗结合后易减弱信号;因此,我们建议不使用含有酪蛋白的封闭剂进行磷酸化特异性抗体的检测。以上针对封闭步骤所提建议均是在以 SignalStain. Boost IHC Detection Reagent 作为检测试剂的情况下提出的。如果选择使用其他检测试剂,我们则建议使用与二抗来源相同的血清进行封闭。抗体:Phospho–Histone H2A.X (Ser139) (20E3) Rabbit mAb #9718样本:石蜡
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