Recombinant
RabMAb

Recombinant Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146)

Reviews (2)Q&A (1)References (16)

Overview

  • Product name

    Anti-Cdk2 (phospho Y15) antibody [EPR2233Y]
    See all Cdk2 primary antibodies

  • Description

    Rabbit monoclonal [EPR2233Y] to Cdk2 (phospho Y15)

  • Host species

    Rabbit

  • Specificity

    ab76146 detects Cdk1, 2, 3, 5 phosphorylated on tyrosine 15

  • Tested applications

    Suitable for: IHC-P, Dot blot, WB, IPmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Cdk2 (phospho Y15). The exact sequence is proprietary.

  • Positive control

    • WB: HeLa cell and mouse spleen tissue lysates; 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged blank expression vector, whole cell lysate; 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK1/2/3/5 expression vector, whole cell lysate IHC-P: Human transitional cell carcinoma of bladder and rat colon tissues. IP: HeLa whole cell lysate (ab150035).

  • General notes

    A trial size is available to purchase for this antibody.

     

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab76146 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
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocol.
Dot blot 1/1000.
WB 1/10000. Detects a band of approximately 33 kDa (predicted molecular weight: 34 kDa).

For unpurified use at 1/1000 - 1/10000.

A band of 65 kDa is also observed in 293T cells overexpressing cdk1, 2, 3, 5 (phospho Y15)
IP 1/100.

For unpurified use at 1/50.
  • Application notes
    Is unsuitable for ICC/IF.

    • Target

    • Function

      Involved in the control of the cell cycle. Interacts with cyclins A, B1, B3, D, or E. Activity of CDK2 is maximal during S phase and G2.

    • Sequence similarities

      Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
      Contains 1 protein kinase domain.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • Cdc2 related protein kinase antibody
      • cdc2-related protein kinase antibody
      • CDC28 antibody
      • CDC2A antibody
      • Cdk 2 antibody
      • CDK1 antibody
      • CDK2 antibody
      • CDK2_HUMAN antibody
      • CDKN2 antibody
      • Cell devision kinase 2 antibody
      • Cell division protein kinase 2 antibody
      • Cyclin dependent kinase 2 antibody
      • cyclin dependent kinase 2-alpha antibody
      • Cyclin-dependent kinase 2 antibody
      • kinase Cdc2 antibody
      • MPF antibody
      • p33 protein kinase antibody
      • p33(CDK2) antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat colon tissue labelling Cdk2 (phospho Y15) with purified ab76146 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9. A prediluted HRP polymer-conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Western blot - Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146)
      Western blot - Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146)
      All lanes : Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146) at 1/1000 dilution

      Lane 1 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged blank expression vector, whole cell lysate
      Lane 2 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK1 expression vector, whole cell lysate
      Lane 3 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK1 expression vector, whole cell lysate. Then the membrane was incubated with phosphatase
      Lane 4 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK2 expression vector, whole cell lysate
      Lane 5 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK2 expression vector, whole cell lysate. Then the membrane was incubated with phosphatase
      Lane 6 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK3 expression vector, whole cell lysate
      Lane 7 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK3 expression vector, whole cell lysate. Then the membrane was incubated with phosphatase
      Lane 8 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK5 expression vector, whole cell lysate
      Lane 9 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK5 expression vector, whole cell lysate. Then the membrane was incubated with phosphatase

      Lysates/proteins at 20 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: 34,65 kDa
      why is the actual band size different from the predicted?



      ab76146 detects Cdk1, 2, 3, 5 phosphorylated on tyrosine 15.

    • Western blot - Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146)
      Western blot - Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146)
      All lanes : Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146) at 1/50000 dilution (purified)

      Lane 1 : Untreated HeLa cell lysate
      Lane 2 : HeLa cell lysate - treated with alkaline phosphatase

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 34 kDa


      Exposure time: 10 seconds


      Blocking and dilution buffer: 5% NFDM/TBST.

    • Dot Blot - Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146)
      Dot Blot - Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146)

      Dot blot analysis of Cdk2 (pY15) peptide (Lane 1) and Cdk2 non-phospho peptide (Lane 2) labelling Cdk2 (pY15) with purified ab76146 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Immunoprecipitation - Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146)
      Immunoprecipitation - Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146)

      ab76146 (purified) at 1/100 immunoprecipitating Cdk2 (phospho Y15) in HeLa cell lysates (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk2 (phospho Y15) antibody [EPR2233Y] (ab76146)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human transitional cell carcinoma of bladder tissue labelling Cdk2 (phospho Y15) with purified ab76146 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9. A prediluted HRP polymer-conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    References

    This product has been referenced in:

    • Qi W  et al. Inhibition of Wee1 sensitizes AML cells to ATR inhibitor VE-822-induced DNA damage and apoptosis. Biochem Pharmacol 164:273-282 (2019). Read more (PubMed: 31014753) »
    • Lim EG  et al. Ethanol extract from Cnidium monnieri (L.) Cusson induces cell cycle arrest and apoptosis via regulation of the p53-independent pathway in HepG2 and Hep3B hepatocellular carcinoma cells. Mol Med Rep 17:2572-2580 (2018). Read more (PubMed: 29207130) »
    See all 16 Publications for this product

    Publishing research using ab76146? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-2 of 2 Abreviews

    Western blot abreview for Anti-Cdk2 (phospho Y15) antibody [EPR2233Y]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    10 µg
    Gel Running Conditions
    Reduced Denaturing (10 %)
    Sample
    Human Cell lysate - whole cell (Hela cells)
    Specification
    Hela cells
    Treatment
    2 ´M MK-1775
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Remi Buisson

    Verified customer

    Submitted Oct 27 2014

    Western blot abreview for Anti-Cdk2 (phospho Y15) antibody [EPR2233Y]

     Excellent
    Abreviews
    abreview image
    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 CDK2, 48h
    Blocking step
    Milk as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 21°C
    Read More

    Dr. Sonia Rocha

    Verified customer

    Submitted Sep 26 2014

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