Product nameAnti-Mad2L1 antibody [17D10]
See all Mad2L1 primary antibodies
DescriptionMouse monoclonal [17D10] to Mad2L1
SpecificityWestern blot with a variety of human cell lines confirms that the antibody is specific for Mad2.
Tested applicationsSuitable for: WB, IP, IHC-Pmore details
Unsuitable for: ICC/IF
Species reactivityReacts with: Human
Recombinant human Mad2.
- WB: HAP1, HeLa and A431 cell lysates
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.1% Sodium azide
Concentration information loading...
PurityProtein A purified
Purification notesPurified from supernatant.
Our Abpromise guarantee covers the use of ab10691 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 2 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 24.6 kDa).|
|IP||Use a concentration of 5 µg/ml.|
|IHC-P||Use at an assay dependent concentration.|
FunctionComponent of the spindle-assembly checkpoint that prevents the onset of anaphase until all chromosomes are properly aligned at the metaphase plate. Required for the execution of the mitotic checkpoint which monitors the process of kinetochore-spindle attachment and inhibits the activity of the anaphase promoting complex by sequestering CDC20 until all chromosomes are aligned at the metaphase plate.
Sequence similaritiesBelongs to the MAD2 family.
Contains 1 HORMA domain.
DomainThe protein has two highly different native conformations, an inactive open conformation that cannot bind CDC20 and that predominates in cytosolic monomers, and an active closed conformation. The protein in the closed conformation preferentially dimerizes with another molecule in the open conformation, but can also form a dimer with a molecule in the closed conformation. Formation of a heterotetrameric core complex containing two molecules of MAD1L1 and of MAD2L1 in the closed conformation promotes binding of another molecule of MAD2L1 in the open conformation and the conversion of the open to the closed form, and thereby promotes interaction with CDC20.
modificationsPhosphorylated on multiple serine residues. The level of phosphorylation varies during the cell cycle and is highest during mitosis. Phosphorylation abolishes interaction with MAD1L1 and reduces interaction with CDC20.
Cellular localizationNucleus. Chromosome > centromere > kinetochore. Cytoplasm. Recruited by MAD1L1 to unattached kinetochores (Probable). Recruited to the nuclear pore complex by TPR during interphase. Recruited to kinetochores in late prometaphase after BUB1, CENPF, BUB1B and CENPE. Kinetochore association requires the presence of NEK2. Kinetochore association is repressed by UBD.
- Information by UniProt
- HsMAD 2 antibody
- HsMAD2 antibody
- MAD 2 antibody
All lanes : Anti-Mad2L1 antibody [17D10] (ab10691) at 2 µg/ml
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Mad2L1 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : A431 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 24.6 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?
Lanes 1 - 4: Merged signal (red and green). Green - ab10691 observed at 24 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab10691 was shown to specifically react with in wild-type HAP1 cells as signal was lost in MAD2L1 knockout cells. Wild-type and MAD2L1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab10691 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 2 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
ab10691 staining Mad2L1 in colon cancer tissue, after antigen retreival, by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Western blot using ab10691 at 1/500.
Lane 1: Hela Nuclear
Lane 2: Hela
Lane 3: A431
Lane 4: Jurkat
Lane 5: 293
Mad2L1 predicted size: 24.6kD.
Secondary ab: Rabbit polyclonal to Mouse IgG H&L (HRP) ab6728 (1/5000)
Exposure time: 1min
20µg of lysate per lane.
Western blot using ab10691 at 1/500. Lane 1: Hela Nuclear Lane 2: Hela Lane 3: A431 Lane 4: Jurkat Lane 5: 293 Mad2L1 predicted size: 24.6kD. Secondary ab: anti-mouse HRP (ab6728) (1/5000) Exposure time: 1min 20µg of lysate per lane.
Mad2L1 antibody (ab10691) at 1/1000 (in TBS for 90 minutes at 22°C) + whole tissue lysate of Human liver biopsy (chronic hepatitis C - (HepC) or non-viral (NVH)) patients at 50µg.
Secondary Antibody: An HRP-conjugated Goat anti-mouse polyclonal (1/100000).
Blocking Step: 5% Milk for 16 hours at 4°C
This product has been referenced in:
- Alfieri C et al. Mechanism for remodelling of the cell cycle checkpoint protein MAD2 by the ATPase TRIP13. Nature 559:274-278 (2018). Read more (PubMed: 29973720) »
- Tambe M et al. Novel Mad2-targeting miR-493-3p controls mitotic fidelity and cancer cells' sensitivity to paclitaxel. Oncotarget 7:12267-85 (2016). WB . Read more (PubMed: 26943585) »