概述

  • 产品名称Anti-MAD2抗体
    参阅全部 MAD2 一抗
  • 描述
    兔多克隆抗体to MAD2
  • 经测试应用适用于: ICC/IF, IHC-P, WB, IPmore details
  • 种属反应性
    与反应: Human
    预测可用于: Guinea pig, Cow, Pig, Chimpanzee, Ferret, Baboon, Rhesus monkey, Gorilla, Orangutan
  • 免疫原

    Synthetic peptide mapping to a region between residues 100 and 150 of human MAD2 (according to SwissProt entry Q13257)

  • 阳性对照
    • HeLa nuclear lysate

性能

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应用

Our Abpromise guarantee covers the use of ab70383 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/1000 - 1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/1000. Detects a band of approximately 24 kDa (predicted molecular weight: 24 kDa).
IP Use at 2-4 µg/mg of lysate.

靶标

  • 功能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.
  • 序列相似性Belongs to the MAD2 family.
    Contains 1 HORMA domain.
  • 结构域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.
  • 翻译后修饰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.
  • 细胞定位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.
  • Information by UniProt
  • 数据库链接
  • 别名
    • HsMAD2 antibody
    • MAD 2 antibody
    • MAD2 like 1 antibody
    • MAD2 mitotic arrest deficient like 1 antibody
    • MAD2-like protein 1 antibody
    • Mad2l1 antibody
    • MD2L1_HUMAN antibody
    • Mitotic arrest deficient 2-like protein 1 antibody
    • Mitotic spindle assembly checkpoint protein MAD2A antibody
    • REV7 antibody
    see all

Anti-MAD2 antibody 图像

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling MAD2 with ab70383 at 1/1000 (1µg/ml). Detection: DAB.
  • Anti-MAD2 antibody (ab70383) at 1 µg/ml + NETN whole cell lysate from HeLa cells at 50 µg
    Developed using the ECL technique

    Predicted band size : 24 kDa
    Observed band size : 24 kDa
    Additional bands at : 38 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 1 minute
  • ab70383 Immunoprecipitating MAD2 in human HeLa whole cell lysate. 200µg of cell lysate was incubated with primary antibody (2 µg/ml in NETN buffer) and matrix (Protein A/G) for 16 hours at 4°C. For western blotting ab70383 (1/1000) was used to confirm successful immunoprecipation.

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  • ICC/IF image of ab70383 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab70383, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Anti-MAD2 antibody (ab70383)参考文献

This product has been referenced in:
  • Eliezer Y  et al. Interplay between the DNA damage proteins MDC1 and ATM in the regulation of the spindle assembly checkpoint. J Biol Chem 289:8182-93 (2014). WB, ICC/IF . Read more (PubMed: 24509855) »
  • Rajanala K  et al. Phosphorylation of nucleoporin Tpr governs its differential localization and is required for its mitotic function. J Cell Sci 127:3505-20 (2014). Read more (PubMed: 24938596) »

See all 3 Publications for this product

Product Wall

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 20 µg
Specification HeLa cells
Gel Running Conditions Reduced Denaturing
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Verified customer

提交于 Sep 23 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (HeLa cells)
Total protein in input 200 µg
Specification HeLa cells
Immuno-precipitation step Protein A/G
Username

Abcam user community

Verified customer

提交于 Sep 21 2010

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