NIH3T3 fibroblast lysates, spiked with 1 ng active GST tagged-TTK protein [Invitrogen, Prod # 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 (Prod # 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.
|Tested species reactivity||Human|
|Published species reactivity||Tag, Rat, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human TTK that contains threonine 33 and serine 37.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Protein kinases are enzymes that transfer a phosphate group from a phosphate donor, generally the g phosphate of ATP, onto an acceptor amino acid in a substrate protein. By this basic mechanism, protein kinases mediate most of the signal transduction in eukaryotic cells, regulating cellular metabolism, transcription, cell cycle progression, cytoskeletal rearrangement and cell movement, apoptosis, and differentiation. With more than 500 gene products, the protein kinase family is one of the largest families of proteins in eukaryotes. The family has been classified in 8 major groups based on sequence comparison of their tyrosine (PTK) or serine/threonine (STK) kinase catalytic domains. The STE group (homologs of yeast Sterile 7, 11, 20 kinases) consists of 50 kinases related to the mitogen-activated protein kinase (MAPK) cascade families (Ste7/MAP2K, Ste11/MAP3K, and Ste20/MAP4K). MAP kinase cascades, consisting of a MAPK and one or more upstream regulatory kinases (MAPKKs) have been best characterized in the yeast pheromone response pathway. Pheromones bind to Ste cell surface receptors and activate yeast MAPK pathway.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Mitotic Checkpoint Kinase Mps1 Has a Role in Normal Physiology which Impacts Clinical Utility.
44-1325G was used in immunocytochemistry to assess Mps1 kinase inhibitors as cancer treatments.
|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||PloS one (10:null)||2015|
Naturally Occurring Mutations in the MPS1 Gene Predispose Cells to Kinase Inhibitor Drug Resistance.
44-1325G was used in immunocytochemistry to identify mutations in the kinase domain of MPS1 that confer resistance against multiple inhibitors.
|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||Cancer research (75:3340)||2015|
||Structure-based design of orally bioavailable 1H-pyrrolo[3,2-c]pyridine inhibitors of mitotic kinase monopolar spindle 1 (MPS1).||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||Journal of medicinal chemistry (56:10045)||2013|
|Structure-based design of orally bioavailable 1H-pyrrolo[3,2-c]pyridine inhibitors of mitotic kinase monopolar spindle 1 (MPS1).||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||Journal of medicinal chemistry (56:10045)||2013|