Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E161] to CDK1 + Cdk2 + Cdk3 (phospho T14)
  • Suitable for: WB, IHC-P, IP, ICC/IF
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

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

  • Description

    Rabbit monoclonal [E161] to CDK1 + Cdk2 + Cdk3 (phospho T14)

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human CDK1 aa 1-100. The exact sequence is proprietary.
    Database link: P06493

  • Positive control

    • WB: HeLa cell lysate. IHC-P: Human lymphoma.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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.

Properties

Images

  • Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    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
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 34 kDa
    Observed band size: 62, 34 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    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.
  • Immunocytochemistry/ Immunofluorescence - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    Immunocytochemistry/ Immunofluorescence - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

    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.

  • Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    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
    All lanes : 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 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    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.
  • Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    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

  • Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    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
    All lanes : 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

  • Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    Western blot - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    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
    All lanes : 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

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

    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 Detection Reagent (ab131366) was used for detection (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK1 + Cdk2 + Cdk3 (phospho T14) antibody [E161] (ab32384)

    Unpurified ab32384, at a 1/50 dilution, staining Cdc2 in paraffin embedded human lymphoma tissue sections by Immunohistochemistry.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

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

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