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Rabbit抗原名称
Phospho-4E-BP1 (Thr37/46)分子别名
Eukaryotic translation initiation factor 4E-binding protein 1; eIF4E-binding protein 1; Phosphorylated heat- and acid-stable protein regulated by insulin 1 (PHAS-I); EIF4EBP1免疫原
Synthetic Peptide细胞定位
Cytoplasm, NucleusAccession
Q13541克隆号
S-744-46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
WB
稀释度
应用 稀释度 推荐种属 WB 1:1000 Hu, Ms, Rt
Phospho-4E-BP1 (Thr37/46) refers to the specific modified form of eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) that has been phosphorylated at its threonine 37 and threonine 46 sites (Thr37/Thr46). 4E-BP1 is a key downstream effector protein of the mTORC1 signaling pathway, and its normal function is to act as a translation repressor: when in a hypophosphorylated state, it tightly binds to eukaryotic translation initiation factor 4E (eIF4E), preventing eIF4E from forming the initiation complex with eIF4G, thereby inhibiting the initiation of cap-dependent translation. However, when cells receive pro-growth signals such as growth factors, nutrients, or energy, the mTORC1 pathway is activated and preferentially catalyzes the phosphorylation of the Thr37 and Thr46 sites on the 4E-BP1 protein. This step is a foundational event for the "hyperphosphorylation" of 4E-BP1; it triggers a cascade of conformational changes in the protein, ultimately causing its dissociation from eIF4E. This release of eIF4E allows for the initiation of ribosome recruitment and protein synthesis. Therefore, detecting the level of Phospho-4E-BP1 (Thr37/46) serves as a highly sensitive and reliable molecular marker for gauging mTORC1 pathway activity. It is widely used as a key indicator in cancer research, metabolic diseases, and the study of cellular growth regulation to assess a cell's translational potential and proliferative status.
免疫印迹
WB result of Phospho-4E-BP1 (Thr37/46) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody: Phospho-4E-BP1 (Thr37/46) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: MCF7 serum starved overnight, then amino acids starved for 1 hour whole cell lysate 6 µg
Lane 2: MCF7 serum starved overnight, then amino acids starved for 1 hour, followed by adding back amino acids for 1 hour and treating with 100 nM insulin for 30 minutes whole cell lysate 6 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 13 kDa
Observed MW: 15-20 kDaWB result of Phospho-4E-BP1 (Thr37/46) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody: Phospho-4E-BP1 (Thr37/46) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: Neuro-2a whole cell lysate 20 µg
Lane 2: C2C12 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 13 kDa
Observed MW: 15-20 kDaWB result of Phospho-4E-BP1 (Thr37/46) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody: Phospho-4E-BP1 (Thr37/46) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: PC-12 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 13 kDa
Observed MW: 15-20 kDa
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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