#5143 MYPT1 Antibody Sampler Kit
|Phospho-MYPT1 (Ser507) Antibody #3040||20 µl||WB||H, M, R||Mk||140||Rabbit|
|Phospho-MYPT1 (Ser668) Antibody #3048||20 µl||WB||H||M, R, Mk||140||Rabbit|
|Phospho-MYPT1 (Thr853) Antibody #4563||20 µl||WB||H, M, R, Hm, Mk, Dg||140||Rabbit|
|Phospho-MYPT1 (Thr696) Antibody #5163||20 µl||WB||H, M, R, Mk||140||Rabbit|
|MYPT1 (D6C1) Rabbit mAb #8574||20 µl||WB, IP, IF-IC||H, Mk, Dg||140||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||WB||Goat|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key: H=Human, M=Mouse, R=Rat, Hm=Hamster, Mk=Monkey, Dg=Dog
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
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 various cell lines using MYPT1 (D6C1) Rabbit mAb.
Western blot analysis of extracts from various cell types using Phospho-MYPT1 (Thr853) Antibody.
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).
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).
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 C2C12 cells, untreated or treated with with the ROCK inhibitor Y27632 using Phospho-MYPT1 (Thr853) Antibody (upper) or MYPT1 Antibody #2634 (lower).
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).
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: 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 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).
Phospho-MYPT1 (Ser507) and phospho-MYPT1 (Ser668) antibodies are directed at sites that were identified at Cell Signaling Technology (CST) using PhosphoScan®, CST's LC-MS/MS platform for modification site discovery. Phosphorylation at these sites was discovered using an Akt substrate antibody. Please visit PhosphoSitePlus®, CST's modification site knowledgebase, at www.phosphosite.org for more information.
DRAQ5 is a registered trademark of Biostatus Limited.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.