| 货号 | 5143T |
| 描述 | The MYPT1 Antibody Sampler Kit is an economical way to examine the total protein levels of MYPT1 as well as MYPT1 phosphorylated at Ser507, Ser668, Thr853, and Thr696. The kit includes enough primary and secondary antibodies to perform four Western blot experiments.MYPT1 Antibody Sampler Kit是一个经济方法去检测MYPT1总蛋白和在Ser507、Ser668、Thr853和Thr696磷酸化的MYPT1蛋白。该试剂盒含有充足的一抗和二抗去进行四次免疫荧光实验。 |
| 目标/特异性 | Each antibody in the MYPT1 Antibody Sampler Kit detects endogenous levels of its target protein. Activation state antibodies detect only target proteins phosphorylated at indicated residues. |
| 供应商 | CST |
| 背景 | Protein phosphatase 1 (PP1) is a ubiquitous eukaryotic protein serine/threonine phosphatase involved in the regulation of various cell functions. Substrate specificity is determined by the binding of a regulatory subunit to the PP1 catalytic subunit (PP1c). It is estimated that over fifty different regulatory subunits exist (1). The myosin phosphatase holoenzyme is composed of three subunits: the PP1 catalytic subunit (PP1c), a targeting/regulatory subunit (MYPT/myosin-binding subunit of myosin phosphatase) and a 20 kDa subunit of unknown function (M20). MYPT binding to PP1cδ alters the conformation of the catalytic cleft and increases enzyme activity and specificity (2). Two MYPT isoforms that are 61% identical have been described. MYPT1 is widely expressed while MYPT2 expression appears to be exclusive to heart and brain (3). Related family members also include MBS85, MYPT3 and TIMAP (4). Myosin phosphatase regulates the interaction of actin and myosin in response to signaling through the small GTPase Rho. Rho activity inhibits myosin phosphatase via Rho-associated kinase (ROCK). Phosphorylation of MYPT1 at Thr696 and Thr853 results in phosphatase inhibition and cytoskeletal reorganization (5,6).Protein phosphatase 1 (PP1)是一个普遍存在的真核蛋白质丝氨酸/苏氨酸磷酸酶，其涉及不同细胞功能的调节。底物的特异性通过一个调节亚单位与PP1 catalytic subunit (PP1c)的结合被确定。据估计大约有五十个不同的调节亚单位存在(1)。myosin 磷酸酶全酶有三个亚单位组成：PP1催化亚单位(PP1c) 、一个靶向/调节亚单位（MYPT/myosin-binding subunit of myosin phosphatase）和一个未知功能的20 kDa大小的亚单位 (M20)。MYPT蛋白结合到PP1cδ蛋白可改变催化部位的构象以及子女国家酶活性和特异性 (2)。61%序列相同的两个MYPT亚单位已经被描述。MYPT1蛋白广泛表达，而MYPT2 蛋白的表达只表达在心脏和大脑组织(3)。相关的家族成员也诱导MBS85、MYPT3和TIMAP (4)。在小GTPase Rho的信号下，Myosin磷酸酶调节actin和myosin相互作用。通过Rho-associated kinase (ROCK)作用，Rho活性抑制myosin磷酸酶。MYPT1蛋白在Thr696和Thr853位点的磷酸化导致磷酸酶抑制和细胞骨架重组(5,6)。 |
| 存放说明 | -20C |
| 参考文献 | Cohen, P.T. (2002) J Cell Sci 115, 241-56. Terrak, M. et al. (2004) Nature 429, 780-4. Fujioka, M. et al. (1998) Genomics 49, 59-68. Ito, M. et al. (2004) Mol Cell Biochem 259, 197-209. Birukova, A.A. et al. (2004) Microvasc Res 67, 64-77. Birukova, A.A. et al. (2004) J Cell Physiol 201, 55-70. |
Western blot analysis of extracts from C2C12 cells, untreated or treated with with the ROCK inhibitor Y27632 using Phospho-MYPT1 (Thr853) Antibody (upper) or MYPT1 Antibody #2634 (lower). | |
Immunoprecipitation of MYPT1 from HeLa cell extracts using MYPT1 (D6C1) Rabbit mAb (lane 2). Western blot was performed using the same antibody. Lane 1 is 10% input. | |
Western blot analysis of extracts from various cell types using Phospho-MYPT1 (Thr853) Antibody. | |
Western blot analysis of extracts from various cell lines using MYPT1 (D6C1) Rabbit mAb. | |
Confocal immunofluorescent analysis of Hep G2 cells using MYPT1 (D6C1) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). | |
Western blot analysis of extracts from 293 cells, untreated or treated with the ROCK inhibitor Y27632 (2 minutes at 10 μM) using Phospho-MYPT1 (Thr696) Antibody #5163 or total MYPT1 Antibody #2634. Phospho-MYPT1 (Thr696) Antibody was pre-incubated as indicated with phospho-MYPT1 (Thr696) peptide or nonphospho-MYPT1 (Thr696) peptide.使用Phospho-MYPT1 (Thr696) Antibody #5163或总的MYPT1 Antibody #2634，免疫印迹（Western Blot）分析293细胞中Phospho-MYPT1 (Ser696)和MYPT1蛋白水平，细胞分为未处理组或Calyculin A #9902处理组。Phospho-MYPT1 (Thr696) Antibody事先孵育phospho-MYPT1 (Thr696) peptide或nonphospho-MYPT1 (Thr696) peptide。 | |
Western blot analysis of extracts from HeLa or NIH/3T3 cells, untreated or treated with the ROCK inhibitor Y-27632, using Phospho-MYPT1 (Thr853) Antibody #4563.使用Phospho-MYPT1 (Thr853) Antibody #4563，免疫印迹（Western Blot）分析HeLa和NIH/3T3细胞中Phospho-MYPT1 (Thr853)蛋白水平，细胞分为未处理和ROCK inhibitor Y-27632处理。 | |
Western blot analysis of extracts from 293 cells, untreated or treated with the ROCK inhibitor Y27632 (2 minutes at 10 μM) using Phospho-MYPT1 (Thr696) Antibody or total MYPT1 Antibody #2634. Phospho-MYPT1 (Thr696) Antibody was pre-incubated as indicated with phospho-MYPT1 (Thr696) peptide or nonphospho-MYPT1 (Thr696) peptide. | |
Western blot analysis of extracts from HeLa and Jurkat cells, untreated or treated with Calyculin A #9902, using Phospho-MYPT1 (Ser668) Antibody #3048 (upper) or MYPT1 Antibody #2634 (lower).使用Phospho-MYPT1 (Ser668) Antibody #3048 (上图)或MYPT1 Antibody #2634 (下图)，免疫印迹（Western Blot）分析HeLa和Jurkat细胞中Phospho-MYPT1 (Ser668)和MYPT1蛋白水平，细胞分为未处理和Calyculin A #9902处理。 | |
Western blot analysis of extracts from HeLa and Jurkat cells, untreated or treated with Calyculin A #9902, using Phospho-MYPT1 (Ser507) Antibody #3040 (upper) or MYPT1 Antibody #2634 (lower).使用Phospho-MYPT1 (Ser507) Antibody #3040 (上图)或MYPT1 Antibody #2634 (下图)，免疫印迹（Western Blot）分析HeLa和Jurkat细胞中Phospho-MYPT1 (Ser507)和MYPT1蛋白水平，细胞分为未处理和Calyculin A #9902处理。 | |
Western blot analysis of extracts from various cell types using MYPT1 Antibody #2634.使用MYPT1 Antibody #2634，免疫印迹（Western Blot）分析不同细胞中MYPT1蛋白水平。 | |
Western blot analysis of extracts from HeLa cells, untreated or treated with Calyculin A #9902, using Phospho-MYPT1 (Ser668) Antibody. The phospho-specificity of the antibody was verified by peptide blocking using no peptide (left), phospho-Ser507 peptide (middle) or phospho-Ser668 peptide (right). | |
Western blot analysis of extracts from HeLa and Jurkat cells, untreated or treated with Calyculin A #9902, using Phospho-MYPT1 (Ser668) Antibody (upper) or total MYPT1 Antibody #2634 (lower). | |
Western blot analysis of extracts from HeLa and Jurkat cells, untreated or treated with calyculin A #9902, using Phospho-MYPT1 (Ser507) Antibody (upper) or MYPT1 Antibody #2634 (lower). | |
Western blot analysis of extracts from HeLa cells, untreated or treated with Calyculin A #9902, using Phospho-MYPT1 (Ser507) Antibody. The phospho-specificity of the antibody was verified by peptide blocking using no peptide (left), phospho-Ser507 peptide (middle) or phospho-Ser668 peptide (right). |
