Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

MYPT1 Antibody Sampler Kit #5143

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-MYPT1 (Ser507) Antibody #3040 40 µl W H M R (Mk) 140 Rabbit
Phospho-MYPT1 (Ser668) Antibody #3048 40 µl W H (M) (R) (Mk) 140 Rabbit
Phospho-MYPT1 (Thr853) Antibody #4563 40 µl W H M R Hm Mk Dg 140 Rabbit
Phospho-MYPT1 (Thr696) Antibody #5163 40 µl W H M R Mk 140 kDa Rabbit
MYPT1 Antibody #2634 40 µl W H M R Hm Mk (C) (Z) (B) (Dg) (Pg) 140 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  C=Chicken  Z=Zebrafish  B=Bovine  Dg=Dog  Pg=Pig
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

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. The MYPT1 Antibody is not likely to cross-react with MYPT2 nor does it cross-react with other family members.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using MYPT1 Antibody #2634.

Western Blotting

Western Blotting

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).

Western Blotting

Western Blotting

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).


Western Blotting

Western Blotting

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.

Western Blotting

Western Blotting

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.

Description

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.

Source / Purification

Phospho-specific polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser507, Ser668, Thr853, and Thr696 of human MYPT1. MYPT1 Antibody is produced by immunizing animals with a synthetic peptide corresponding to amino-terminal residues of human MYPT1. Antibodies are purified using protein A and peptide affinity chromatography.

Background

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.

  1. Cohen, P.T. (2002) J Cell Sci 115, 241-56.
  2. Terrak, M. et al. (2004) Nature 429, 780-4.
  3. Fujioka, M. et al. (1998) Genomics 49, 59-68.
  4. Ito, M. et al. (2004) Mol Cell Biochem 259, 197-209.
  5. Birukova, A.A. et al. (2004) Microvasc Res 67, 64-77.
  6. Birukova, A.A. et al. (2004) J Cell Physiol 201, 55-70.

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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