The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/5000 - 1/20000. Detects a band of approximately 24 kDa (predicted molecular weight: 24 kDa).
Use at 2-4 µg/mg of lysate.
Component 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.
Belongs to the MAD2 family. Contains 1 HORMA domain.
The 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.
Phosphorylated 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. Phosphorylated by NEK2.
Nucleus. Chromosome > centromere > kinetochore. Cytoplasm. Cytoplasm > cytoskeleton > spindle pole. 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.
Detection of Human MAD2 by Immunoprecipitation in NETN whole cell lysate from HeLa cells (600 µg/IP) using ab70385 at 0.7 µg/mg for IP and at 0.2 µg/ml for subsequent WB detection. Note: bands between 50 and 60 kDa represent the IgG chains.
Tan CL et al. Stranglehold on the spindle assembly checkpoint: the human papillomavirus E2 protein provokes BUBR1-dependent aneuploidy. Cell Cycle14:1459-70 (2015).
Read more (PubMed: 25789401) »
Jahn SC et al. Constitutive Cdk2 activity promotes aneuploidy while altering the spindle assembly and tetraploidy checkpoints. J Cell Sci126:1207-17 (2013).
Read more (PubMed: 23321641) »