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5143
MYPT1 Antibody Sampler Kit

MYPT1 Antibody Sampler Kit #5143

Western Blotting Image 1

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).

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Western Blotting Image 2

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).

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Western Blotting Image 3

Western blot analysis of extracts from various cell types using Phospho-MYPT1 (Thr853) Antibody.

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Western Blotting Image 4

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.

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Western Blotting Image 5

Western blot analysis of extracts from various cell lines using MYPT1 (D6C1) Rabbit mAb.

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Western Blotting Image 6

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.

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Western Blotting Image 7

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).

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Western Blotting Image 8

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).

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Western Blotting Image 9

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).

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IP Image 10

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.

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IF-IC Image 11

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).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-MYPT1 (Ser507) Antibody 3040 20 µl
  • WB
H M R 140 Rabbit 
Phospho-MYPT1 (Ser668) Antibody 3048 20 µl
  • WB
H 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
H Mk Dg 140 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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 two Western blot experiments.

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.

Phospho-specific polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser507, Ser668, Thr853, and Thr696 of human MYPT1. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human MYPT1 portein.

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.
Entrez-Gene Id
4659
Swiss-Prot Acc.
O14974
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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