Anti-TTK/Mps1 antibody [N1] - BSA free (ab11108)
Key features and details
- Mouse monoclonal [N1] to TTK/Mps1 - BSA free
- Suitable for: Flow Cyt, ICC/IF, IP, WB
- Reacts with: Human, Recombinant fragment
- Isotype: IgG1
Overview
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Product name
Anti-TTK/Mps1 antibody [N1] - BSA free
See all TTK/Mps1 primary antibodies -
Description
Mouse monoclonal [N1] to TTK/Mps1 - BSA free -
Host species
Mouse -
Tested applications
Suitable for: Flow Cyt, ICC/IF, IP, WBmore details -
Species reactivity
Reacts with: Human, Recombinant fragment -
Immunogen
Recombinant fragment within Human TTK/Mps1 aa 1-900. The exact sequence is proprietary. (catalytically inactive - D663A)
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Epitope
This antibody recognizes an epitope in the N-terminal region of Mps1. -
Positive control
- In Flow Cytometry, this antibody gave a positive signal in methanol fixed/Tween permeabilised HeLa cells.
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General notes
This antibody clone is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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Purity
IgG fraction -
Clonality
Monoclonal -
Clone number
N1 -
Myeloma
PAIB3Ag81 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab11108 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt | Use 2µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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ICC/IF | Use a concentration of 1 µg/ml. | |
IP | Use at an assay dependent concentration. | |
WB | Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 95 kDa). NOT SUITABLE for blocking with BSA. Block in 5% milk for 1 hour. |
Target
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Function
Phosphorylates proteins on serine, threonine, and tyrosine. Probably associated with cell proliferation. Essential for chromosome alignment by enhancing AURKB activity (via direct CDCA8 phosphorylation) at the centromere, and for the mitotic checkpoint. -
Tissue specificity
Present in rapidly proliferating cell lines. -
Sequence similarities
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family.
Contains 1 protein kinase domain. - Information by UniProt
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Database links
- Entrez Gene: 7272 Human
- Omim: 604092 Human
- SwissProt: P33981 Human
- Unigene: 169840 Human
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Alternative names
- cancer/testis antigen 96 antibody
- CT96 antibody
- Dual specificity protein kinase TTK antibody
see all
Images
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All lanes : Anti-TTK/Mps1 antibody [N1] - BSA free (ab11108) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 4 : Jurkat nuclear extract lysate (ab14844)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 95 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Exposure time: 8 minutes
NOTE: MILK was used to block non-specific binding, rather than BSA. -
TTK/Mps1 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Mouse monoclonal to Mps1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab11108.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band: 100kDa: TTK/Mps1 -
ICC/IF image of ab11108 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab11108, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
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Overlay histogram showing HeLa cells stained with ab11108 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab11108, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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Anti-TTK/Mps1 antibody [N1] - BSA free (ab11108) at 10 µg/ml +
Recombinant human TTK/Mps1 protein (ab89589) at 0.01 µg
Secondary
Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 95 kDa
Exposure time: 8 minutesab11108 recognizes the full length tagged recombinant TTK/Mps1 protein (ab89589) which has an expected molecular weight of 130 kDa.
Protocols
References (17)
ab11108 has been referenced in 17 publications.
- Gui P et al. Mps1 dimerization and multisite interactions with Ndc80 complex enable responsive spindle assembly checkpoint signaling. J Mol Cell Biol 12:486-498 (2020). PubMed: 32219319
- Raghavendra A et al. Expression of MAGE-A and NY-ESO-1 cancer/testis antigens is enriched in triple-negative invasive breast cancers. Histopathology 73:68-80 (2018). PubMed: 29465777
- Choi M et al. TC Mps1 12, a novel Mps1 inhibitor, suppresses the growth of hepatocellular carcinoma cells via the accumulation of chromosomal instability. Br J Pharmacol 174:1810-1825 (2017). PubMed: 28299790
- Zhang J et al. Downregulation of tyrosine threonine kinase inhibits tumor growth via G2/M arrest in human endometrioid endometrial adenocarcinoma. Tumour Biol 39:1010428317712444 (2017). PubMed: 28718377
- Wild T et al. The Spindle Assembly Checkpoint Is Not Essential for Viability of Human Cells with Genetically Lowered APC/C Activity. Cell Rep 14:1829-40 (2016). WB ; Human . PubMed: 26904940