Anti-Mps1 antibody [N1] - BSA free (ab11108)

Overview

  • Product nameAnti-Mps1 antibody [N1] - BSA free
    See all Mps1 primary antibodies
  • Description
    Mouse monoclonal [N1] to Mps1 - BSA free
  • SpecificityThis antibody was shown to specifically detect Mps1 in Stucke et al. 2002.
  • Tested applicationsSuitable for: Flow Cyt, ICC/IF, Dot blot, Functional Studies, IP, ELISA, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant near full length protein (catalytically inactive - D663A), corresponding to amino acids 3-856 of Human Mps1.

  • EpitopeThis 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.
  • General notesWe can conjugate this antibody to FITC for you (please see ab150213 for details).

Properties

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
Flow Cyt Use 2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC/IF Use a concentration of 1 µg/ml.
Dot blot Use at an assay dependent concentration.
Functional Studies Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ELISA 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

  • FunctionPhosphorylates 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 specificityPresent in rapidly proliferating cell lines.
  • Sequence similaritiesBelongs to the protein kinase superfamily. Ser/Thr protein kinase family.
    Contains 1 protein kinase domain.
  • Information by UniProt
  • Database links
  • Alternative names
    • cancer/testis antigen 96 antibody
    • CT96 antibody
    • Dual specificity protein kinase TTK antibody
    • ECSTY kinase antibody
    • ESK antibody
    • FLJ38280 antibody
    • hMPS1 antibody
    • Monopolar Spindle 1 Like 1 antibody
    • MPH1 antibody
    • Mps 1 antibody
    • MPS1L1 antibody
    • Phosphotyrosine Picked Threonine Kinase antibody
    • Phosphotyrosine picked threonine-protein kinase antibody
    • PYT antibody
    • RP3-357D13.2 antibody
    • ttk antibody
    • TTK Protein Kinase antibody
    • TTK_HUMAN antibody
    see all

Anti-Mps1 antibody [N1] - BSA free images

  • All lanes : Anti-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
    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.
  • 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: 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).

  • 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.

References for Anti-Mps1 antibody [N1] - BSA free (ab11108)

This product has been referenced in:
  • 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 . Read more (PubMed: 26904940) »
  • Bailey ML  et al. Dependence of Human Colorectal Cells Lacking the FBW7 Tumor Suppressor on the Spindle Assembly Checkpoint. Genetics 201:885-95 (2015). WB ; Human . Read more (PubMed: 26354767) »

See all 10 Publications for this product

Product Wall

Application Western blot
Loading amount 50 µg
Gel Running Conditions Non-reduced Denaturing (8)
Sample Human Cell lysate - whole cell (Human colon carcinoma cells)
Specification Human colon carcinoma cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Eleni Petsalaki

Verified customer

Submitted Mar 11 2014

Application Western blot
Loading amount 50 µg
Gel Running Conditions Non-reduced Denaturing (8)
Sample Hamster Cell lysate - whole cell (Chinese Hamster Ovary cells)
Specification Chinese Hamster Ovary cells
Blocking step (agent) for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Eleni Petsalaki

Verified customer

Submitted Feb 27 2014

Application Immunoprecipitation
Immuno-precipitation step Protein A/G
Sample Hamster Cell lysate - whole cell (Chinese Hamster Ovary cells)
Specification Chinese Hamster Ovary cells
Total protein in input 50 µg
Username

Dr. Eleni Petsalaki

Verified customer

Submitted Feb 27 2014

Application Immunoprecipitation
Immuno-precipitation step Protein A/G
Sample Human Cell lysate - whole cell (Human colon carcinoma cells)
Specification Human colon carcinoma cells
Total protein in input 50 µg
Username

Dr. Eleni Petsalaki

Verified customer

Submitted Feb 27 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa)
Loading amount 10 µg
Specification HeLa
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 15°C
Username

Ms. AeLee Jeong

Verified customer

Submitted Apr 12 2013

Thank you for your email.



Lab has confirmed that the recommended concentration is actually 1ug/ml; the 10ug/ml has been added in error which will be corrected soon. With the new recommended dilutions, this antibody will be suitable...

Read More
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Methanol
Permeabilization No
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Ms. Chelsea Backer

Verified customer

Submitted Apr 14 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa cells)
Loading amount 10 µg
Specification HeLa cells
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 27°C
Username

Abcam user community

Verified customer

Submitted Mar 23 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2%
Username

Abcam user community

Verified customer

Submitted Aug 08 2005

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"