Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)

Reviews (1) Submit a question References (3)
Western blot - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)
  • Western blot - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)
  • Western blot - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)
  • Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4513-54-3] to Cdk4
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Cdk4 antibody [EPR4513-54-3]
    See all Cdk4 primary antibodies

  • Description

    Rabbit monoclonal [EPR4513-54-3] to Cdk4

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IFmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, MCF7 and K562 cell lysates. ICC/IF: Wild-type HAP1 and HeLa cells.

  • 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.

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

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108355 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)
1/1000 - 1/10000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
ICC/IF
1/400.

This product gave a positive signal in HAP1 cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min)
Notes
WB
1/1000 - 1/10000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
ICC/IF
1/400.

This product gave a positive signal in HAP1 cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min)
Application notes
Is unsuitable for IHC-P.

Target

  • Function

    Ser/Thr-kinase component of cyclin D-CDK4 (DC) complexes that phosphorylate and inhibit members of the retinoblastoma (RB) protein family including RB1 and regulate the cell-cycle during G(1)/S transition. Phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complexes and the subsequent transcription of E2F target genes which are responsible for the progression through the G(1) phase. Hypophosphorylates RB1 in early G(1) phase. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. Also phosphorylates SMAD3 in a cell-cycle-dependent manner and represses its transcriptional activity. Component of the ternary complex, cyclin D/CDK4/CDKN1B, required for nuclear translocation and activity of the cyclin D-CDK4 complex.

  • Involvement in disease

    Defects in CDK4 are a cause of susceptibility to cutaneous malignant melanoma type 3 (CMM3) [MIM:609048]. Malignant melanoma is a malignant neoplasm of melanocytes, arising de novo or from a pre-existing benign nevus, which occurs most often in the skin but also may involve other sites.

  • Sequence similarities

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

  • Post-translational
    modifications

    Phosphorylation at Thr-172 is required for enzymatic activity. Phosphorylated, in vitro, at this site by CCNH-CDK7, but, in vivo, appears to be phosphorylated by a proline-directed kinase. In the cyclin D-CDK4-CDKN1B complex, this phosphorylation and consequent CDK4 enzyme activity, is dependent on the tyrosine phosphorylation state of CDKN1B. Thus, in proliferating cells, CDK4 within the complex is phosphorylated on Thr-172 in the T-loop. In resting cells, phosphorylation on Thr-172 is prevented by the non-tyrosine-phosphorylated form of CDKN1B.

  • Cellular localization

    Cytoplasm. Nucleus. Membrane. Cytoplasmic when non-complexed. Forms a cyclin D-CDK4 complex in the cytoplasm as cells progress through G(1) phase. The complex accumulates on the nuclear membrane and enters the nucleus on transition from G(1) to S phase. Also present in nucleoli and heterochromatin lumps. Colocalizes with RB1 after release into the nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • Cdk 4 antibody
      • cdk4 antibody
      • CDK4 protein antibody
      • CDK4_HUMAN antibody
      • Cell division kinase 4 antibody
      • Cell division protein kinase 4 antibody
      • CMM 3 antibody
      • CMM3 antibody
      • Crk3 antibody
      • Cyclin dependent kinase 4 antibody
      • Cyclin-dependent kinase 4 antibody
      • Melanoma cutaneous malignant 3 antibody
      • MGC14458 antibody
      • p34 cdk4 antibody
      • PSK J3 antibody
      • PSK-J3 antibody
      see all

    Images

    • Western blot - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)
      Western blot - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)

      Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
      Lanes 2, 4 and 6: CDK4 knockout HAP1 cell lysate (20 µg)
      Lanes 1 and 2: Green signal from target - ab108355 observed at 34 kDa
      Lanes 3 and 4: Red signal from loading control - ab8226 observed at 42 kDa
      Lanes 5 and 6: Merged (red and green) signal
      ab108355 was shown to specifically react with CDK4 when CDK4 knockout samples were used. Wild-type and CDK4 knockout samples were subjected to SDS-PAGE. ab108355 and ab8226 (loading control to beta actin) were both diluted at 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)
      Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)

      ab108355 staining Cdk4 in wild-type HAP1 cells (top panel) and Cdk4 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab108355 at 1/400 dilution and ab7291 at 1μg/ml concentration overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150120) (shown in pseudo colour red). Nuclear DNA was labelled in blue with DAPI.

    • Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)
      Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)

      Immunofluorescence staining of MCF7 cells with purified ab108355 at a working dilution of 1/100, 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 ab108355 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-Cdk4 antibody [EPR4513-54-3] (ab108355)
      Western blot - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)
      All lanes : Anti-Cdk4 antibody [EPR4513-54-3] (ab108355) at 1/5000 dilution (purified)

      Lane 1 : HeLa cell lysate
      Lane 2 : K562 cell lysate
      Lane 3 : Ramos cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

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



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

    • Western blot - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)
      Western blot - Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)This image is courtesy of an Abreview submitted by Sonia Rocha
      All lanes : Anti-Cdk4 antibody [EPR4513-54-3] (ab108355) at 1/1000 dilution (unpurified)

      Lane 1 : Human osteosarcoma cells - control, non-targeting siRNA
      Lane 2 : Human osteosarcoma cells - siRNA for CDK4

      Lysates/proteins at 20 µg per lane.

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

      Developed using the ECL technique.

      Performed under reducing conditions.

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


      Exposure time: 5 seconds

      See Abreview

    • Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)
      Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)

    References (3)

    Publishing research using ab108355? Please let us know so that we can cite the reference in this datasheet.

    ab108355 has been referenced in 3 publications.

    • Pinelli S  et al. Overexpression of microRNA-486 affects the proliferation and chemosensitivity of mesothelioma cell lines by targeting PIM1. Int J Mol Med 47:N/A (2021). PubMed: 33955505
    • Xu K  et al. LncRNA FOXD2-AS1 plays an oncogenic role in hepatocellular carcinoma through epigenetically silencing CDKN1B(p27) via EZH2. Exp Cell Res 380:198-204 (2019). PubMed: 31004581
    • Weng TY  et al. Homoharringtonine induced immune alteration for an Efficient Anti-tumor Response in Mouse Models of Non-small Cell Lung Adenocarcinoma Expressing Kras Mutation. Sci Rep 8:8216 (2018). PubMed: 29844447

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Western blot abreview for Anti-Cdk4 antibody [EPR4513]

     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 CDK4, 48h
    Blocking step
    Milk as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 21°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Sonia Rocha

    Verified customer

    Submitted Sep 26 2014

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com