Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

Overview

  • Product nameAnti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161]
  • Description
    Rabbit monoclonal [E161] to CDK1 + Cdk2 + Cdk3 (phospho T14)
  • Tested applicationsSuitable for: WB, IHC-P, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human CDK1 aa 1-100.
    Database link: P06493

  • Positive control
    • WB: HeLa cell lysate. IHC-P: Human lymphoma.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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.

    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.

    A trial size is available to purchase for this antibody.

     

    Alternative versions available:

    Anti-CDK1 antibody (Alexa Fluor® 488) [E161] (ab193829)

    Anti-CDK1 antibody (HRP) [E161] (ab193834)

Properties

Applications

Our Abpromise guarantee covers the use of ab32384 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 1/1000 - 1/10000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
IHC-P 1/50 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol)

IP 1/50.
ICC/IF 1/100 - 1/500.

Target

Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] images

  • All lanes : Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384) at 1/2000 dilution (purified)

    Lane 1 : Empty vector transfected 293T (human embryonic kidney) whole cell lysates
    Lane 2 : CDK1-GFP transfected 293T (human embryonic kidney) whole cell lysates
    Lane 3 : CDK2-GFP transfected 293T (human embryonic kidney) whole cell lysates
    Lane 4 : CDK3-GFP transfected 293T (human embryonic kidney) whole cell lysates
    Lane 5 : CDK5-GFP transfected 293T (human embryonic kidney) whole cell lysates

    Lysates/proteins at 15 µg per lane.

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

    Predicted band size : 34 kDa
    Observed band size : kDa (why is the actual band size different from the predicted?)


    Exposure time : 15 seconds

    Blocking/Diluting buffer 5% NFDM/TBST

    34kDa band represents endogenous CDK, and 62kDa represents CDK-GFP.

  • Immunohistochemical staining of paraffin embedded human B cell lymphoma with purified ab32384 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is 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.
  • Immunofluorescence staining of HeLa cells with purified ab32384 at a working dilution of 1/200, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab32384 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • All lanes : Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384) at 1/2000 dilution (purified)

    Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 3mM hydroxyurea for 20 hours whole cell lysates
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 3mM hydroxyurea for 20 hours whole cell lysates. Then the membrane was incubated with phosphatase.

    Lysates/proteins at 15 µg per lane.

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

    Predicted band size : 34 kDa
    Observed band size : 34 kDa


    Exposure time : 30 seconds

    Blocking/Diluting buffer 

  • Immunohistochemical staining of paraffin embedded human ovarian carcinoma with purified ab32384 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is 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.
  • Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384) at 1/5000 dilution (purified) + Raw264.7 cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size : 34 kDa
    Observed band size : 34 kDa

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • All lanes : Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384) at 1/10000 dilution (purified)

    Lane 1 : C6 cell lysate
    Lane 2 : PC-12 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size : 34 kDa
    Observed band size : 34 kDa

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • All lanes : Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384) at 1/10000 dilution (purified)

    Lane 1 : A431 cell lysate
    Lane 2 : HepG2 cell lysate
    Lane 3 : SK-BR-3 cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size : 34 kDa
    Observed band size : 34 kDa

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • ab32384 (purified) at 1/50 immunoprecipitating CDK1 in 10 μg HEK293 (Lanes 1 and 2, observed at 34 kDa). Lane 3 - PBS. For western blotting, HRP Veriblot for IP (ab131366) was used as the secondary antibody (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
  • Unpurified ab32384, at a 1/50 dilution, staining Cdc2 in paraffin embedded human lymphoma tissue sections by Immunohistochemistry.

  • All lanes : Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384) at 1/1000 dilution (unpurified)

    Lane 1 : A431 cell lysate
    Lane 2 : HepG2 cell lysate
    Lane 3 : SKBR3 cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.


    Predicted band size : 34 kDa

References for Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

This product has been referenced in:
  • He Y  et al. Cell Division Cycle 6 Promotes Mitotic Slippage and Contributes to Drug Resistance in Paclitaxel-Treated Cancer Cells. PLoS One 11:e0162633 (2016). WB ; Human . Read more (PubMed: 27611665) »
  • Wang J  et al. Prognostic significance of G2/M arrest signaling pathway proteins in advanced non-small cell lung cancer patients. Oncol Lett 9:1266-1272 (2015). IHC ; Mouse . Read more (PubMed: 25663895) »

See all 6 Publications for this product

Product Wall

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Human Cell lysate - whole cell (osteosarcoma cells)
Specification osteosarcoma cells
Treatment lane 1 control, lane 2 siRNA for CDK1, 48h
Blocking step Milk as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 21°C
Username

Dr. Sonia Rocha

Verified customer

Submitted Sep 26 2014

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