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  1. Link

    cdk4-antibody-epr2513y-ab68266.pdf

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Cell Biology Cell Cycle Kinases/Phosphatases Cdks
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RecombinantRabMAb

Recombinant Anti-Cdk4 antibody [EPR2513Y] (ab68266)

  • Datasheet
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Western blot - Anti-Cdk4 antibody [EPR2513Y] (ab68266)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR2513Y] (ab68266)
  • Immunoprecipitation - Anti-Cdk4 antibody [EPR2513Y] (ab68266)
  • Flow Cytometry (Intracellular) - Anti-Cdk4 antibody [EPR2513Y] (ab68266)
  • Anti-Cdk4 antibody [EPR2513Y] (ab68266)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2513Y] to Cdk4
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Carrier Free PE

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Recombinant Human Cdk4 protein (ab126909)
Secondary
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Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

View more associated products

Overview

  • Product name

    Anti-Cdk4 antibody [EPR2513Y]
    See all Cdk4 primary antibodies
  • Description

    Rabbit monoclonal [EPR2513Y] to Cdk4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, WB, IPmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human Cdk4 aa 1-100 (N terminal).

  • Positive control

    • WB: HeLa, Ramos, and K562 cell lysates. Rat and mouse lung tissue lysates. ICC/IF: MCF7 cells. Flow Cyt (intra) (Intra): A549 cells. IP: K562 whole cell lysate.
  • 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR2513Y
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Kinases/Phosphatases
    • Cdks
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Kinases/Phosphatases
    • Cdks
    • Cancer
    • Cell cycle
    • Kinases/phosphatases
    • Cdks
    • Isotype/Loading Controls
    • Loading Controls
    • Cdk4

Associated products

  • Alternative Versions

    • PE Anti-Cdk4 antibody [EPR2513Y] (ab213664)
    • Alexa Fluor® 488 Anti-Cdk4 antibody [EPR2513Y] (ab214642)
    • Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • K-562 nuclear extract lysate (ab14851)
    • MCF7 nuclear extract lysate (ab14860)
  • Recombinant Protein

    • Recombinant Human Cdk4 protein (ab126909)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab68266 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
Flow Cyt (Intra)
1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/100.
WB
1/500 - 1/1000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
IP
1/30.
Notes
Flow Cyt (Intra)
1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/100.
WB
1/500 - 1/1000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
IP
1/30.
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.
  • Target information above from: UniProt accession P11802 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1019 Human
    • Entrez Gene: 12567 Mouse
    • Entrez Gene: 94201 Rat
    • Omim: 123829 Human
    • SwissProt: P11802 Human
    • SwissProt: P30285 Mouse
    • SwissProt: P35426 Rat
    • Unigene: 95577 Human
    • Unigene: 6839 Mouse
    • Unigene: 6115 Rat
    see all
  • 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 [EPR2513Y] (ab68266)
    Western blot - Anti-Cdk4 antibody [EPR2513Y] (ab68266)
    All lanes : Anti-Cdk4 antibody [EPR2513Y] (ab68266) at 1/2000 dilution

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : K562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
    Lane 3 : Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate
    Lane 4 : Rat lung lysate
    Lane 5 : Mouse lung lysate

    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
    Additional bands at: 34 kDa. We are unsure as to the identity of these extra bands.



    Exposure times:  Lane 1-3: 30 secs; Lane 4-5: 180 secs.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR2513Y] (ab68266)
    Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR2513Y] (ab68266)

    Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cdk4 with ab68266 at 1:100 (5.4 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% triton X-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain.

    PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunoprecipitation - Anti-Cdk4 antibody [EPR2513Y] (ab68266)
    Immunoprecipitation - Anti-Cdk4 antibody [EPR2513Y] (ab68266)

    ab68266 at 1:30 dilution (2 µg) immunoprecipitating Cdk4 in K562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate.
    Lane 1 (input): K562 whole cell lysate 10 µg
    Lane 2 (+): ab68266 & K562 whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab68266 in K562 whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Flow Cytometry (Intracellular) - Anti-Cdk4 antibody [EPR2513Y] (ab68266)
    Flow Cytometry (Intracellular) - Anti-Cdk4 antibody [EPR2513Y] (ab68266)

    Intracellular Flow Cytometry analysis of A549 (Human lung carcinoma epithelial cell) cells labeling Cdk4 with ab68266 at 1/50 dilution (1µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). 

     

     

  • Anti-Cdk4 antibody [EPR2513Y] (ab68266)
    Anti-Cdk4 antibody [EPR2513Y] (ab68266)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (16)

Publishing research using ab68266? Please let us know so that we can cite the reference in this datasheet.

ab68266 has been referenced in 16 publications.

  • Lv T  et al. Knockdown of lncRNA DLEU1 inhibits the tumorigenesis of oral squamous cell carcinoma via regulation of miR-149-5p/CDK6 axis. Mol Med Rep 23:N/A (2021). PubMed: 33880596
  • Zhang Y  et al. LncRNA-CCDC144NL-AS1 Promotes the Development of Hepatocellular Carcinoma by Inducing WDR5 Expression via Sponging miR-940. J Hepatocell Carcinoma 8:333-348 (2021). PubMed: 33977095
  • Stegmann KM  et al. The folate antagonist methotrexate diminishes replication of the coronavirus SARS-CoV-2 and enhances the antiviral efficacy of remdesivir in cell culture models. Virus Res 302:198469 (2021). PubMed: 34090962
  • Baptista Moreno Martin AC  et al. [10]-Gingerol improves doxorubicin anticancer activity and decreases its side effects in triple negative breast cancer models. Cell Oncol (Dordr) 43:915-929 (2020). PubMed: 32761561
  • Wei L  et al. TMPO-AS1, a Novel E2F1-Regulated lncRNA, Contributes to the Proliferation of Lung Adenocarcinoma Cells via Modulating miR-326/SOX12 Axis. Cancer Manag Res 12:12403-12414 (2020). PubMed: 33293866
View all Publications for this product

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