|Application ||WB, IP|
|Calculated MW||23510 Da|
|Application & Usage||Western blotting (1:500 – 1:2000) and Immunoprecipitation. HeLa cell lysate can be used as a positive control. However, the optimal concentrations should be determined individually. The antibody recognizes the MAD2 of human origin. Reactivity to other species has not been tested. Reactivity with mouse samples is likely due to high sequence homology.|
|Other Names||MAD2, HsMAD2, Mitotic Arrest Deficient 2, MAD2L1, MAD2-like 1|
|Formulation||100 µl affinity purified rabbit polyclonal antibody in phosphate-buffered saline (PBS) containing 30% glycerol, 1% BSA and 0.02% thimerosal.|
|Handling||The antibody solution should be gently mixed before use.|
|Reconstitution & Storage||-20 °C|
|Precautions||MAD2 Antibody is for research use only and not for use in diagnostic or therapeutic procedures.|
|Function||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.|
|Cellular Location||Nucleus. Chromosome, centromere, kinetochore. Cytoplasm. Cytoplasm, cytoskeleton, spindle pole Note=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.|
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Provided below are standard protocols that you may find useful for product applications.
Cell cycle progression is subject to arrest at the mitotic spindle assembly checkpoint in response to incorrect spindle fiber assembly. MAD2 (for mitotic arrest-deficient) is a component of the mitotic spindle checkpoint. Cells with mutated MAD2 do not undergo mitotic arrest in response to incorrect spindle fiber assembly, which results in missegregation and eventual cell death. A breast carcinoma cell line with reduced MAD2 expression, T47D, was shown to complete mitosis in the presence of nocodazole, an inhibitor of mitotic spindle assembly. MAD2 is localized to unattached kinetochores during prometaphase and disassociates upon spindle fiber attachment, indicating that MAD2 regulates kinetochore binding to the spindle fibers. Human MAD2 has also been shown to associate with insulin receptor (IR), but not IGFIR, implicating MAD2 as a mediator for IR-specific signaling. MAD2B, a MAD2 homolog, is required for the execution of the mitotic checkpoint monitoring the kinetochore-spindle attachment process and if the process is not complete, MAD2B delays the onset of anaphase.
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