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Phospho-TTK (Thr33, Ser37) Polyclonal Antibody

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Phospho-TTK (Thr33, Ser37) Antibody in Western Blot (WB) — NIH3T3 fibroblast lysates, spiked with 1 ng active GST tagged-TTK protein (Invitrogen, Product # PV3792) (2-6) or Kinase dead mutant TTK protein (1) were resolved on a 10% polyacrylamide gel and transferred to PVDF. The membrane was blocked with a 3% BSA-TBST buffer for one hour at room temperature, and then incubated with the TTK (pTpS33/37) antibody for two hours at room temperature in 3% BSA-TBST buffer, following prior incubation with: no peptide (2), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), a generic phosphothreonine-containing peptide (4), a generic phosphoserine (5), or the phosphopeptide immunogen corresponding to TTK (pTpS33/37) (6). After washing, the membrane was incubated with goat F (ab')2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and signals were detected using the Pierce SuperSignal™ reagent. The data show that the signal detected in the active TTK protein was selectively blocked by the phosphopeptide corresponding to TTK (pTpS33/37). The data also show the loss of phosphorylation signal on kinase dead protein indicating that the signal is phosphorylation site-specific.

Phospho-TTK (Thr33, Ser37) Antibody in Immunoprecipitation (IP) — Figure 1 CCT271850 inhibits MPS1 autophosphorylation in cells. ( A ) Structure of CCT271850. ( B ) HCT116 cells transiently expressing Myc-tagged MPS1 were treated with increasing concentrations of CCT271850 for 2 h. Cell lysates were analysed for MPS1 autophosphorylation (at pT33pS37 and pT676 sites) and gel shift using immunoblotting. ( C ) HCT116 cells were synchronised with nocodazole for 24 h followed by treatment with indicated concentrations of CCT271850 for 2 h in the presence of MG132. Cell lysates were immunoprecipitated for endogenous MPS1 with phospho-specific anti-pT33pS37 antibodies and immunoblotted with anti-MPS1 antibodies. Total cell lysates were also immunoblotted with anti-MPS1 antibodies as a control for immunoprecipitation input (lower panel). ( D ) Immunofluorescence for MPS1 pT33pS37 localisation at kinetochores with and without treatment with CCT271850. Cells were treated with CCT271850 for 1 h, followed by treatment with nocodazole, MG132 and CCT271850 for an additional hour. Cells were fixed and stained as indicated.

Product Details

44-1325G

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000	-
Immunocytochemistry (ICC/IF)	-	View 4 publications 4 publications
ELISA (ELISA)	-	View 1 publication 1 publication
Immunoprecipitation (IP)	Assay-dependent	View 1 publication 1 publication
Miscellaneous PubMed (Misc)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human
Published species	Human, Rat, Tag
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human TTK that contains threonine 33 and serine 37. View immunogen
Conjugate	Unconjugated
Form	Liquid
Purification	Antigen affinity chromatography
Storage buffer	Dulbecco's PBS, pH 7.3, with 1mg/mL BSA, 50% glycerol
Contains	0.05% sodium azide
Storage conditions	-20°C
RRID	AB_2533594

Target Information

This gene encodes a constant regulatory subunit of protein phosphatase 2. Protein phosphatase 2 is one of the four major Ser/Thr phosphatases, and it is implicated in the negative control of cell growth and division. It consists of a common heteromeric core enzyme, which is composed of a catalytic subunit and a constant regulatory subunit, that associates with a variety of regulatory subunits. The constant regulatory subunit A serves as a scaffolding molecule to coordinate the assembly of the catalytic subunit and a variable regulatory B subunit. This gene encodes an alpha isoform of the constant regulatory subunit A. Alternatively spliced transcript variants have been described.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

A Biosensor for the Mitotic Kinase MPS1 Reveals Spatiotemporal Activity Dynamics and Regulation.

Current biology : CB

Kuijt TEF,Lambers MLA,Weterings S,Ponsioen B,Bolhaqueiro ACF,Staijen DHM,Kops GJPL

44-1325G was used in Immunocytochemistry to develop MPS1sen, a sensitive and specific FRET-based biosensor for MPS1 activity o understand when and where MPS1 activation occurs and how MPS1 signaling is modulated during mitosis.

Mon Oct 05 00:00:00 EDT 2020

Checkpoint signaling and error correction require regulation of the MPS1 T-loop by PP2A-B56.

The Journal of cell biology

Hayward D,Bancroft J,Mangat D,Alfonso-Pérez T,Dugdale S,McCarthy J,Barr FA,Gruneberg U

44-1325G was used in Immunocytochemistry-immunoflourescence to investigate if the BUBR1-bound pool of PP2A-B56 regulates monopolar spindle 1 T-loop autophosphorylation and hence its activation status in mammalian cells.

Mon Oct 07 00:00:00 EDT 2019

Characterisation of CCT271850, a selective, oral and potent MPS1 inhibitor, used to directly measure in vivo MPS1 inhibition vs therapeutic efficacy.

British journal of cancer

Faisal A,Mak GWY,Gurden MD,Xavier CPR,Anderhub SJ,Innocenti P,Westwood IM,Naud S,Hayes A,Box G,Valenti MR,De Haven Brandon AK,O'Fee L,Schmitt J,Woodward HL,Burke R,vanMontfort RLM,Blagg J,Raynaud FI,Eccles SA,Hoelder S,Linardopoulos S

Published figure using Phospho-TTK (Thr33, Ser37) polyclonal antibody (Product # 44-1325G) in Immunoprecipitation

Tue Apr 25 00:00:00 EDT 2017

Mitotic Checkpoint Kinase Mps1 Has a Role in Normal Physiology which Impacts Clinical Utility.

PloS one

Martinez R,Blasina A,Hallin JF,Hu W,Rymer I,Fan J,Hoffman RL,Murphy S,Marx M,Yanochko G,Trajkovic D,Dinh D,Timofeevski S,Zhu Z,Sun P,Lappin PB,Murray BW

44-1325G was used in immunocytochemistry to assess Mps1 kinase inhibitors as cancer treatments.

Tue Jun 14 00:00:00 EDT 2016

Naturally Occurring Mutations in the MPS1 Gene Predispose Cells to Kinase Inhibitor Drug Resistance.

Cancer research

Gurden MD,Westwood IM,Faisal A,Naud S,Cheung KM,McAndrew C,Wood A,Schmitt J,Boxall K,Mak G,Workman P,Burke R,Hoelder S,Blagg J,Van Montfort RL,Linardopoulos S

44-1325G was used in immunocytochemistry to identify mutations in the kinase domain of MPS1 that confer resistance against multiple inhibitors.

Sat Aug 15 00:00:00 EDT 2015

Structure-based design of orally bioavailable 1H-pyrrolo[3,2-c]pyridine inhibitors of mitotic kinase monopolar spindle 1 (MPS1).

Journal of medicinal chemistry

Naud S,Westwood IM,Faisal A,Sheldrake P,Bavetsias V,Atrash B,Cheung KM,Liu M,Hayes A,Schmitt J,Wood A,Choi V,Boxall K,Mak G,Gurden M,Valenti M,de Haven Brandon A,Henley A,Baker R,McAndrew C,Matijssen B,Burke R,Hoelder S,Eccles SA,Raynaud FI,Linardopoulos S,van Montfort RL,Blagg J

Fri Dec 27 00:00:00 EST 2013

Structure-based design of orally bioavailable 1H-pyrrolo[3,2-c]pyridine inhibitors of mitotic kinase monopolar spindle 1 (MPS1).

Journal of medicinal chemistry

Fri Dec 27 00:00:00 EST 2013

Structure-based design of orally bioavailable 1H-pyrrolo[3,2-c]pyridine inhibitors of mitotic kinase monopolar spindle 1 (MPS1).

Journal of medicinal chemistry

Fri Dec 27 00:00:00 EST 2013

Bioinformatics

Protein Aliases: cancer/testis antigen 96; Dual specificity protein kinase TTK; monopolar spindle 1 kinase; monopolar spindle 1-like 1; OTTHUMP00000016778; OTTHUMP00000220743; phosphotyrosine picked threonine kinase; Phosphotyrosine picked threonine-protein kinase; PYT

Gene Aliases: CT96; ESK; MPH1; MPS1; MPS1L1; PYT; TTK

UniProt ID: (Human) P33981

Entrez Gene ID: (Human) 7272

Molecular Function: non-receptor serine/threonine protein kinase

Function(s)

spindle organization mitotic spindle organization mitotic spindle assembly checkpoint positive regulation of cell proliferation positive regulation of pathway-restricted SMAD protein phosphorylation peptidyl-tyrosine phosphorylation chromosome separation

Process(es)

protein serine/threonine kinase activity protein serine/threonine/tyrosine kinase activity protein tyrosine kinase activity protein binding ATP binding

Phospho-TTK (Thr33, Ser37) Polyclonal Antibody

Product Details

44-1325G

Western Blot (WB)

Immunocytochemistry (ICC/IF)

ELISA (ELISA)

Immunoprecipitation (IP)

Miscellaneous PubMed (Misc)

Species Reactivity

Published species

Host/Isotype

Class

Type

Immunogen

Conjugate

Form

Purification

Storage buffer

Contains

Storage conditions

RRID

Target Information

Bioinformatics

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