Recombinant
RabMAb

Anti-CDK1+CDK2+CDK5+CDK3 (phospho Y15) antibody [EPR7875] (ab133463)

Overview

  • Product name
    Anti-CDK1+CDK2+CDK5+CDK3 (phospho Y15) antibody [EPR7875]
  • Description
    Rabbit monoclonal [EPR7875] to CDK1+CDK2+CDK5+CDK3 (phospho Y15)
  • Host species
    Rabbit
  • Specificity
    The antibody will cross-react with phosphorylated CDK1 (pY15), CDK2 (pY15), CDK3 (pY15) and CDK5 (pY15) but not with non-phosphorylated CDK1, CDK2, CDK3 and CDK5. Please see our ELISA results on the images section.
  • Tested applications
    Suitable for: WB, IHC-P, ELISA, Dot blotmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human CDK1+CDK2+CDK5+CDK3 (phospho Y15). The exact sequence is proprietary.

  • Positive control
    • HeLa cell lysate treated with UV, human colon, C6 cell lysate, NIH/3T3 cell lysate, Saso2 cells, human breast carcinoma tissue
  • 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 ab133463 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 - 1/2000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
IHC-P 1/50 - 1/75. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ELISA Use at an assay dependent concentration.
Dot blot 1/1000.

Images

  • Anti-CDK1+CDK2+CDK5+CDK3 (phospho Y15) antibody [EPR7875] (ab133463) at 1/1000 dilution (purified) + HeLa cell lysate treated with UV at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded human colon with purified ab133463 at a working dilution of 1 in 75. The secondary antibody used is a HRP polymer for rabbit IgG. 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.

  • Direct ELISA antibody dose-response curve using purified ab133463 at 0-1000 ng/ml. Antigen concentration of 1000 ng/mL. An alkaline phosphatase-conjugated goat anti-rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

  • Direct ELISA antibody dose-response curve using purified ab133463 at 0-1000 ng/ml. Antigen concentration of 1000 ng/mL. An alkaline phosphatase-conjugated goat anti-rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

  • Anti-CDK1+CDK2+CDK5+CDK3 (phospho Y15) antibody [EPR7875] (ab133463) at 1/2000 dilution (purified) + C6 cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Anti-CDK1+CDK2+CDK5+CDK3 (phospho Y15) antibody [EPR7875] (ab133463) at 1/2000 dilution (purified) + NIH/3T3 cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Dot blot analysis of CDK1 (pY15) phospho peptide (lane 1) and CDK1 non-phospho peptide (lane 2) labelling CDK1 (phospho Y15) with unpurified ab133463 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 10 seconds.

  • Saso2 cells were incubated at 37°C for 24 hours with vehicle control (0 μM) and different concentrations of Amifostine (ab141060). Decreased expression of CDK1 (phospho Y15) (unpurified ab133463) in Saso2 cells correlates with an increase in Amifostine concentration, as described in literature.

    Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with unpurified ab133463 at 1 μg/ml and ab32384 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 and visualised using ECL development solution.

  • Immunohistochemical analysis of paraffin embedded human breast carcinoma tissue labelling CDK1 with unpurified ab133463 at 1/50 dilution.

  • Anti-CDK1+CDK2+CDK5+CDK3 (phospho Y15) antibody [EPR7875] (ab133463) at 1/1500 dilution (unpurified) + NIH/3T3 cell lysate at 1/1000 dilution

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Anti-CDK1+CDK2+CDK5+CDK3 (phospho Y15) antibody [EPR7875] (ab133463) at 1/1500 dilution (unpurified) + C6 cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • All lanes : Anti-CDK1+CDK2+CDK5+CDK3 (phospho Y15) antibody [EPR7875] (ab133463) at 1/1000 dilution (unpurified)

    Lane 1 : HeLa cell lysate
    Lane 2 : HeLa cell lysate treated with UV

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit IgG at 1/2000 dilution

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

  • Immunohistochemical staining of paraffin embedded human colon with unpurified ab133463 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. 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.

References

This product has been referenced in:
  • Jin LB  et al. Paeoniflorin induces G2/M cell cycle arrest and caspase-dependent apoptosis through the upregulation of Bcl-2 X-associated protein and downregulation of B-cell lymphoma 2 in human osteosarcoma cells. Mol Med Rep 17:5095-5101 (2018). Read more (PubMed: 29363721) »
  • 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) »
See all 6 Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Permeabilization
Yes - 0.5% Triton X100 in PBS
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Jul 10 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (12,5%)
Sample
Human Purified protein (Human primary CD4 T cells)
Specification
Human primary CD4 T cells
Treatment
TCR activated using anti-CD3/CD28 for 72h and treated with genotoxic agent to induce cell cycle arrest
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Mr. Marco Craveiro

Verified customer

Submitted Jul 23 2013

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

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