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Rabbit抗原名称
Phospho-S6 Ribosomal Protein (Ser235/236)分子别名
Phospho-Small ribosomal subunit protein eS6 (Ser235/236), Phospho-40S ribosomal protein S6 (Ser235/236), Phospho-Phosphoprotein NP33 (Ser235/236)细胞定位
Nucleolus, Cytoplasm, NucleusAccession
P62753克隆号
S-R203抗体类型
Rabbit mAb翻译后修饰类型
磷酸化反应种属 ?
Hu, Ms, Rt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
ICC ?
WB
IP
稀释度
应用 稀释度 推荐种属 WB 1:1000 Hu, Ms, Rt ICC 1:500 Hu IP 1:50 Hu
Ribosomal protein S6 (rpS6 or eS6) is a component of the 40S ribosomal subunit and is therefore involved in translation. Mouse model studies have shown that phosphorylation of eS6 is involved in the regulation of cell size, cell proliferation, and glucose homeostasis [PMID: 16166381, PMID: 16679021, PMID: 19479038]. p70S6K is a serine/threonine kinase that phosphorylates the S6 protein of the 40S ribosomal subunit (phosphorylated S6 ribosomal protein (phospho-S6rp)) at several sites, including serines 235 and 236, leading to initiation of protein synthesis [PMID: 16860938].
免疫印迹
WB result of Phospho-S6 Ribosomal Protein (Ser235/236) Rabbit mAb Primary antibody: Phospho-S6 Ribosomal Protein (Ser235/236) Rabbit mAb at 1/1000 dilution Lane 1: HeLa whole cell lysate 5 µg Lane 2: HeLa serum starvation 16 hour then treated with calyculin (100 nM, 30 min) whole cell lysate 5 µg Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 29 kDa Observed MW: 32 kDa
WB result of Phospho-S6 Ribosomal Protein (Ser235/236) Rabbit mAb Primary antibody: Phospho-S6 Ribosomal Protein (Ser235/236) Rabbit mAb at 1/1000 dilution Lane 1: NIH/3T3 whole cell lysate 5 µg Lane 2: NIH/3T3 treated with CA (100 nM, 30 min) whole cell lysate 5 µg Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 29 kDa Observed MW: 32 kDa
WB result of Phospho-S6 Ribosomal Protein (Ser235/236) Rabbit mAb Primary antibody: Phospho-S6 Ribosomal Protein (Ser235/236) Rabbit mAb at 1/1000 dilution Lane 1: C6 whole cell lysate 5 µg Lane 2: C6 treated with CA (100 nM, 30 min) whole cell lysate 5 µg Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 29 kDa Observed MW: 32 kDa
免疫沉淀
Phospho-S6 Ribosomal Protein (Ser235/236) Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating Phospho-S6 Ribosomal Protein (Ser235/236) in 0.4 mg HeLa serum starvation 16 hour then treated with calyculin (100 nM, 30 min) whole cell lysate.
Western blot was performed on the immunoprecipitate using Phospho-S6 Ribosomal Protein (Ser235/236) Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: HeLa serum starvation 16 hour then treated with calyculin (100 nM, 30 min) whole cell lysate 20 µg (Input)
Lane 2: Phospho-S6 Ribosomal Protein (Ser235/236) Rabbit mAb IP in HeLa serum starvation 16 hour then treated with calyculin (100 nM, 30 min) whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in HeLa serum starvation 16 hour then treated with calyculin (100 nM, 30 min) whole cell lysate
Predicted MW: 29 kDa
Observed MW: 32 kDa
免疫细胞化学
ICC analysis of HeLa cells treated with insulin (150nM,
6mins) (top panel) and HeLa cells treated with Rapamycin (10nM,2h) (below panel). Anti- Phospho-S6 Ribosomal Protein (Ser235/236) 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).
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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