Overview

  • Product name
  • Description
    Rabbit polyclonal to CDCA2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 950 to the C-terminus of Human CDCA2.

    Read Abcam's proprietary immunogen policy (Peptide available as ab45267.)

  • Positive control
    • This antibody gave a positive control in the following human lysates: Hela nuclear HepG2 nuclear Jurkat nuclear (data not shown)

Properties

Applications

Our Abpromise guarantee covers the use of ab45129 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 125 kDa (predicted molecular weight: 113 kDa).
ICC/IF Use a concentration of 5 µg/ml.

Target

  • Relevance
    CDCA2 is a regulator of chromosome structure during mitosis. It is required for condensin-depleted chromosomes to retain their compact architecture through anaphase. It acts by mediating the recruitment of phosphatase PP1-gamma subunit (PPP1CC) to chromatin at anaphase and into the following interphase. At anaphase onset, its association with chromatin targets a pool of PPP1CC to dephosphorylate substrates. CDCA2 is essential for cell viability.
  • Cellular localization
    Nuclear. Excluded from the nucleolus. Present in nucleoplasm throughout the G1, S and G2 stages of the cell cycle. During M phase, it becomes diffuse throughout the cell as the nuclear membrane breaks down, and faintly accumulates later on metaphase chromatin. As the cell progresses to anaphase, it accumulates on chromatin.
  • Database links
  • Alternative names
    • CDCA 2 antibody
    • Cell division cycle associated protein 2 antibody
    • PPP1R81 antibody
    • protein phosphatase 1, regulatory subunit 81 antibody
    • Recruits PP1 onto mitotic chromatin at anaphase protein antibody
    • Repo Man antibody
    see all

Images

  • All lanes : Anti-CDCA2 antibody (ab45129) 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 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : HepG2 nuclear extract lysate (ab14660)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 113 kDa
    Observed band size: 125 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 40 kDa. We are unsure as to the identity of these extra bands.

  • ICC/IF image of ab45129 stained HepG2 cells. The cells were 4% PFA fixed (10 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 (ab45129, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

References

This product has been referenced in:
  • Qian J  et al. Cdk1 orders mitotic events through coordination of a chromosome-associated phosphatase switch. Nat Commun 6:10215 (2015). Read more (PubMed: 26674376) »
  • Wurzenberger C  et al. Sds22 and Repo-Man stabilize chromosome segregation by counteracting Aurora B on anaphase kinetochores. J Cell Biol 198:173-83 (2012). Read more (PubMed: 22801782) »
See all 2 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa, HEK293, U2OS)
Loading amount
30 µg
Specification
HeLa, HEK293, U2OS
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Mr. Antibody Solutions

Verified customer

Submitted Oct 08 2008

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

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