概述

  • 产品名称Anti-Cdc25C抗体[E302]
    参阅全部 Cdc25C 一抗
  • 描述
    兔单克隆抗体[E302] to Cdc25C
  • 特异性The antibody can also detect splice isoform 2, 4 and 5 of human Cdc25C, based on sequence homology.
  • 经测试应用适用于: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    A synthetic peptide corresponding to residues near N-terminus of human Cdc25C.

  • 阳性对照
    • HeLa cell lysate and human urinary bladder carcinoma tissue.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Alternative versions available:

    Anti-Cdc25C antibody (Alexa Fluor® 488) [E302] (ab205425)
    Anti-Cdc25C antibody (Alexa Fluor® 647) [E302] (ab205426)

性能

应用

Our Abpromise guarantee covers the use of ab32444 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
WB 1/1000 - 1/5000. Detects a band of approximately 60 kDa (predicted molecular weight: 53 kDa).
IHC-P 1/2500.

For unpurified, use 1/250 - 1/500.

ICC/IF 1/250 - 1/500.
Flow Cyt 1/180.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/30 - 1/80.

靶标

  • 功能Functions as a dosage-dependent inducer in mitotic control. It is a tyrosine protein phosphatase required for progression of the cell cycle. It directly dephosphorylates CDK1 and activate its kinase activity.
  • 序列相似性Belongs to the MPI phosphatase family.
    Contains 1 rhodanese domain.
  • 发展阶段Expressed predominantly in G2 phase.
  • 翻译后修饰Phosphorylated by CHK1 on Ser-216. This phosphorylation creates a binding site for 14-3-3 protein and inhibits the phosphatase. Phosphorylated by PLK4.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • CDC 25 antibody
    • Cdc 25C antibody
    • CDC25 antibody
    • CDC25C antibody
    • Cell division cycle 25 homolog C antibody
    • Cell division cycle 25C antibody
    • Cell division cycle 25C protein antibody
    • Dual specificity phosphatase Cdc25C antibody
    • M phase inducer phosphatase 3 antibody
    • M-phase inducer phosphatase 3 antibody
    • Mitosis inducer CDC25 antibody
    • MPIP3 antibody
    • MPIP3_HUMAN antibody
    • Phosphotyrosine phosphatase antibody
    • PPP1R60 antibody
    • protein phosphatase 1, regulatory subunit 60 antibody
    see all

Anti-Cdc25C antibody [E302] 图像



  • Predicted band size : 53 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Cdc25C knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: Hu bladder cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab32444 observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab32444 was shown to recognize Cdc25C when Cdc25C knockout samples were used, along with additional cross-reactive bands. Wild-type and Cdc25C knockout samples were subjected to SDS-PAGE. ab32444 and ab8245 (loading control to GAPDH) were diluted at 1/2500 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

  • All lanes : Anti-Cdc25C antibody [E302] (ab32444) at 1/5000 dilution (purified)

    Lane 1 : K562 (human chronic myelogenous leukemia) whole cell lysate
    Lane 2 : HEK293 (human embryonic kidney) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 53 kDa
    Observed band size : 60 kDa (why is the actual band size different from the predicted?)

    Blocking and diluting buffer 5% NFDM/TBST

  • Anti-Cdc25C antibody [E302] (ab32444) at 1/1000 dilution (purified) + HeLa (human cervix adenocarcinoma) whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 53 kDa
    Observed band size : 60 kDa (why is the actual band size different from the predicted?)

    Blocking and diluting buffer 5% NFDM/TBST

  • Immunohistochemical analysis of paraffin embedded human pancreas tissue section labelling Cdc25C with purified ab32444 at dilution of 1/2500. The secondary antibody used was Goat Anti-Rabbit IgG H&L (HRP) (ab97051), at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling Cdc25C with purified ab32444 at 1/400. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000) using ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary. Nuclei couterstained with DAPI (blue).

    For negative control 1, rabbit primary antibody was used, followed by anti-mouse secondary antibody (ab150120). For negative control 2, mouse primary antibody (ab7291) was used followed by anti-rabbit secondary antibody (ab150077). 

  • Anti-Cdc25C antibody [E302] (ab32444) at 1/5000 dilution (unpurified) + HeLa cell lysate

    Predicted band size : 53 kDa
    Observed band size : 60 kDa (why is the actual band size different from the predicted?)
  • Immunohistochemical analysis of paraffin-embedded human urinary bladder carcinoma unpurified ab32444 at 1/250 dilution.

    Immunohistochemical analysis of paraffin-embedded human urinary bladder carcinoma ab32444 at 1/250 dilution.
  • Flow Cytometry analysis of K562 (human chronic myelogenous leukemia) cells labelling Cdc25C with purified ab32444 at 1/180 (red). Cells were fixed with 4% paraformaldehyde. Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Flow cytometry analysis of HeLa cells, staining Cdc25C with unpurified ab32444.

    Cells were fixed with formaldehyde and permeabilized with 90% methanol. Samples were incubated with primary antibody (1/20 in PBS + 10% goat serum) for 1 hour at 23°C. A FITC-conjugated goat anti-rabbit polyclonal IgG (1/1000) was used as the secondary antibody.

    See Abreview

  • Ab32444 (purified) at 1/30 immunoprecipitating Cdc25C in HeLa (human cervix adenocarcinoma) whole cell lysate.

    Lane 1 (input): HeLa (human cervix adenocarcinoma) whole cell lysate

    Lane 2 (+): ab32444 + HeLa (human cervix adenocarcinoma) whole cell lysate

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32444 in HeLa (human cervix adenocarcinoma) whole cell lysate

    For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/10000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

Anti-Cdc25C antibody [E302] (ab32444)参考文献

This product has been referenced in:
  • Chuang MJ  et al. The HDAC inhibitor LBH589 induces ERK-dependent prometaphase arrest in prostate cancer via HDAC6 inactivation and down-regulation. PLoS One 8:e73401 (2013). WB, ICC/IF ; Human . Read more (PubMed: 24023871) »
  • Kimura M  et al. Involvement of multiple cell cycle aberrations in early preneoplastic liver cell lesions by tumor promotion with thioacetamide in a two-stage rat hepatocarcinogenesis model. Exp Toxicol Pathol 65:979-88 (2013). Rat . Read more (PubMed: 23474136) »

See all 4 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 cell lysate)
Loading amount 20 µg
Specification HeLa cell lysate
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 May 15 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Human Fibroblast)
Loading amount 15 µg
Specification Human Fibroblast
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris Nupage gel)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 25 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Human Cell (HeLa Cells)
Specification HeLa Cells
Preparation Cell harvesting/tissue preparation method: Trypsin/EDTA
Sample buffer: Complete Media then PBS
Fixation Formaldehyde
Permeabilization Yes - 90% Methanol
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Dr. Brandon White

Verified customer

提交于 Aug 30 2012

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