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Rabbit抗原名称
Phospho-gamma H2A.X (Ser139)分子别名
Histone H2AX; H2a/x; Histone H2A.X; H2AFX; H2AX免疫原
Synthetic Peptide细胞定位
NucleusAccession
P16104克隆号
S-981-56抗体类型
Recombinant mAb抗体同种型
IgG翻译后修饰类型
磷酸化反应种属 ?
Hu, Ms, Rt阳性样本
HeLa纯化方式
Protein A浓度
0.5 mg/ml标记
Unconjugated性状
Liquid缓冲体系
PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300
储存条件
12 months from date of receipt / reconstitution, -20 °C as supplied
应用
IHC-P ?
ICC ?
WB
IP
稀释度
应用 稀释度 推荐种属 Dot Blot 1:1000 WB 1:1000 Hu IP 1:50 Hu IHC-P 1:200 Hu, Ms, Rt ICC 1:500 Hu
Phospho-gamma H2A.X (Ser139), also known as γ-H2AX, is a histone variant that plays a crucial role in the DNA damage response (DDR). It is phosphorylated at serine 139 by kinases such as ATM and ATR in response to DNA double-strand breaks (DSBs), which are often caused by ionizing radiation or cytotoxic agents. This phosphorylation event is one of the earliest cellular responses to DNA damage and results in the formation of γ-H2AX foci in the cell nucleus, marking the sites of DSBs on a one-to-one basis. These foci serve as recruitment signals for DNA repair proteins, facilitating the repair process and maintaining genomic stability. Due to its sensitivity and specificity, γ-H2AX is widely used as a biomarker for DNA damage and repair in research, including studies on cancer, aging, and environmental genotoxicity.
免疫印迹
WB result of Phospho-gamma H2A.X (Ser139) Recombinant Rabbit mAb
Primary antibody: Phospho-gamma H2A.X (Ser139) gamma Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated HeLa whole cell lysate 20 µg
Lane 2: HeLa treated with 100 ng/ml Nocodazole for 17 hours whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
免疫沉淀
Phospho-gamma H2A.X (Ser139) Recombinant Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating Phospho-gamma H2A.X (Ser139) in 0.4 mg HeLa+Nocodazole(100ng/ml,17h) whole cell lysate.
Western blot was performed on the immunoprecipitate using Phospho-gamma H2A.X (Ser139) Recombinant Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: HeLa +Nocodazole (100ng/ml,17h) whole cell lysate 20 µg (Input)
Lane 2: Phospho-gamma H2A.X (Ser139) Recombinant Rabbit mAb IP in HeLa +Nocodazole (100ng/ml,17h) whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in HeLa + Nocodazole (100ng/ml,17h) whole cell lysate
Predicted MW: 15 kDa
Observed MW: 17 kDa
斑点杂交
Dot blot result of Phospho-gamma H2A.X (Ser139) Recombinant Rabbit mAb
Lane1: gamma H2A.X (Ser139) phospho peptide
Lane2: gamma H2A.X unmodified peptide
Primary antibody: Phospho-gamma H2A.X (Ser139) Recombinant Rabbit mAb at 1/1000 dilution
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
免疫组化
IHC shows positive staining in paraffin-embedded human ovarian cancer (Left) and negative staining in human ovarian cancer treated with phosphatase (Right). Anti- Phospho-gamma H2A.X (Ser139) antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse testis (Left) and negative staining in mouse testis treated with phosphatase (Right). Anti- Phospho-gamma H2A.X (Ser139) antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat spleen (Left) and negative staining in rat spleen treated with phosphatase (Right). Anti- Phospho-gamma H2A.X (Ser139) antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
免疫细胞化学
ICC analysis of HeLa cells exposured to 10 J/㎡UV and then recovered two hours and untreated HeLa cells (below panel). Anti- gamma H2A.X (phospho S139) antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
组织表达图谱
Expression of Phospho-gamma H2A.X (Ser139) in tumor tissue.
Expression of Phospho-gamma H2A.X (Ser139) in human tissue.
Expression of Phospho-gamma H2A.X (Ser139) in mouse & rat tissue.
FAQs
我们一般不推荐客户回收利用抗体。 因为抗体使用之后缓冲体系已经发生改变，不同客户在回收抗体的保存条件上也会有差异，所以抗体回收使用效果无法保证。另外，我们对一批抗体回收验证测试，测试结果显示不同抗体可回收次数不同，一般效价越高的抗体，可重复使用的次数越多，客户可根据实验情况来确定
我们推荐客户使用TPST+5%脱脂奶粉来稀释一抗，进行封闭。 虽然BSA被推荐为WB检测磷酸化蛋白的常用封闭剂，但是脱脂奶粉获取更加方便，覆盖更广泛的非特异性结合位点，在一抗性能优越的前提下，使用脱脂奶粉封闭性价比更高
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