3 天之前 · Polyclonal Antibody for studying AMPK-alpha1 (Thr183) phosphate/AMPK-alpha2 (Thr172) phosphate. Validated for WB. Available in 2 sizes. Highly specific and rigorously validated in-house, Phospho-AMPKα (Thr172) Antibody (CST #2531) is ready to ship.

大家好，我尝试做western blot看pAMPK／AMPK的比值变化，看文献以此评价AMPK系统激活程度。 问题是，pAMPK和AMPK的分子量都是60-62，感觉不能用同一张膜染，但是如果染在两张

AMP激活蛋白激酶 (AMPK)是一种高度保守的蛋白激酶，在调节能量平衡方面发挥重要作用。 AMPK是一个由AMPKα、β和γ亚基形成的异源三聚体，α为催化亚基，β和γ为调节亚基。 3种亚基又存在不同的亚型 (α1和α2，β1和β2，γ1、γ2和γ3)，不同亚型由不同的基因编码。 热休克、缺氧、缺血等导致的应激反应，可引起AMP/ATP比例增高，导致AMPK激活。 AMPK可被上游激酶磷酸化而被激活，如AMPK Kinase (AMPKK)和LKB1。 肿瘤抑制因子LKB1可磷酸化AMPKα …

抗体是CST的兔抗（#5256S）pAMPK THR172位点，应该算公认好用的。 跑过N次了，一般是浓度高的为双条带，浓度低的为单条带，是我蛋白上样量太多了吗？

请各位大侠指教，最近做AMPK和P-AMPK得WB，AMPK和P-AMPK总是做不出来条带 (TUBLINE条带能出来)。 一抗抗体是CELL SIGANL公司的。 二抗用的是anti-mouse800.anti-rat700.最后条带扫描用ODYSSEY.样本是小鼠骨骼肌，用的2%SDS裂解液（10mM Tris-Hcl PH:6.8, 10%SDS, GLYCERIN, H2O),裂解蛋白时加了罗氏的COCKTAIL和氟化钠.电泳先60V，30MIN,后110Vz直到MARKER跑开，转膜用的PVDF膜，100V,60MIN，20%甲醇NO …

Invitrogen Anti-Phospho-AMPK alpha-1,2 (Thr183, Thr172) Polyclonal, Catalog # 44-1150G. Tested in Western Blot (WB), Immunocytochemistry (ICC/IF) and Immunohistochemistry (Paraffin) (IHC (P)) applications. This antibody reacts with Human, Mouse samples. Supplied as 100 µL purified antibody.

WB: 适用于变性蛋白样本的免疫印迹检测. IHC: 适用于组织样本的石蜡 (IHC-p)或冰冻 (IHC-f)切片样本的免疫组化/荧光检测. IF/ICC: 适用于细胞样本的荧光检测. ELISA (peptide): 适用于抗原肽的ELISA检测. 反应: Human,Mouse,Rat 预测:

AMPKα是总量，p-AMPKα是磷酸化的AMPK，做WB时，先孵育p-AMPKα的一抗，测出磷酸化的量，再用stripping buffer洗掉抗体，孵育AMPKα一抗，测定总量，最后用p-AMPKα/AMPKα，得出其磷酸化水平。

Invitrogen Anti-Phospho-AMPK alpha-1,2 (Thr183, Thr172) Polyclonal, Catalog # 44-1150G. Tested in Western Blot (WB), Immunocytochemistry (ICC/IF) and Immunohistochemistry (Paraffin) (IHC (P)) applications. This antibody reacts with Human, Mouse samples. Supplied as 100 µL purified antibody.

Invitrogen Anti-Phospho-AMPK alpha-1,2 (Thr172) Polyclonal, Catalog # PA5-37821. Tested in Western Blot (WB), Immunocytochemistry (ICC/IF) and Immunohistochemistry (Paraffin) (IHC (P)) applications. This antibody reacts with Human, Mouse, Rat samples. Supplied as 100 µL purified antibody (1 mg/mL).