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Rabbit Recombinant Monoclonal CDK4 antibody. Suitable for IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 161 publications.


Images

Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (AB108357), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk4 antibody [EPR4513-32-7] (AB108357), expandable thumbnail
  • Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (AB108357), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-32-7] (AB108357), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-Cdk4 antibody [EPR4513-32-7] (AB108357), expandable thumbnail

Publications

  • Cell death & disease 14:8372023
    Upregulation of FAM83F by c-Myc promotes cervical cancer growth and aerobic glycolysis via Wnt/β-catenin signaling activation.
    Applications:
    Unspecified application
    Reactive species:
    Unspecified reactive species
    Changlin Zhang,Lixiang Liu,Weizhao Li,Mengxiong Li,Xunzhi Zhang,Chi Zhang,Huan Yang,Jiayuan Xie,Wei Pan,Xue Guo,Peng She,Li Zhong,Tian Li
    PubMed 38104106
  • Inflammation 47:421-4372023
    lncRNA HOTAIRM1 Activated by HOXA4 Drives HUVEC Proliferation Through Direct Interaction with Protein Partner HSPA5.
    Applications:
    Unspecified application
    Reactive species:
    Unspecified reactive species
    Yu Zhou,Qiang Wu,Xiangshu Long,Youfu He,Jing Huang
    PubMed 37898994

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested

Tested
Tested

Species

Human

Dilution info

1/100 - 1/250

Notes

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

Tested
Tested

Species

Human

Dilution info

1/1000 - 1/10000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

Notes

-

Tested
Tested

Species

Human

Dilution info

1/10 - 1/100

Notes

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

Associated Products

Select an associated product type

14 products for Alternative Product

5 products for Alternative Version

Target data

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.

Alternative names

Recommended products

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Rabbit Recombinant Monoclonal CDK4 antibody. Suitable for IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 161 publications.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR4513-32-7

Purification technique

Affinity purification Protein A

Dissociation constant

1.86 x 10-11 M

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Stable for 12 months at -20°C

Notes

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

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

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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

  • Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357), expandable thumbnail

    Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357)

    Lanes 1- 2: Merged signal (red and green). Green - ab108357 observed at 34 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.

    ab108357 was shown to react with Cdk4 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human CDK4 knockout HeLa cell line ab255378 (knockout cell lysate Human CDK4 knockout HeLa cell lysate ab263780) was used. Wild-type HeLa and CDK4 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108357 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357) at 1/1000 dilution

    Lane 1: Wild-type HeLa cell lysate at 20 µg

    Lane 2: CDK4 knockout HeLa cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 34 kDa, 55 kDa

    Observed band size: 34 kDa, 55 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labeling Cdk4 with purified ab108357 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer (pH 9). Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L), was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357), expandable thumbnail

    Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357)

    Lanes 5 and 6: Merged signal (red and green). Green - ab108357 observed at 34kDa. Red - loading control to beta actin observed at 40kDa.

    ab108357 was shown to specifically react with CDK4 in wild-type HAP1 cells. No band was observed when CDK4 knockout samples were examined. Wild-type and CDK4 knockout samples were subjected to SDS-PAGE. ab108357 and Anti-beta Actin antibody [mAbcam 8226] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    Lanes 1 - 4: Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357) at 1/1000 dilution

    Lanes 5 - 6: Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357)

    Lanes 1, 3 and 5: Wild-type HAP1 cell lysate at 20 µg

    Lanes 2, 4 and 6: CDK4 knockout HAP1 cell lysate at 20 µg

    Predicted band size: 34 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357)

    Immunocytochemical analysis of MCF7 (human breast adenocarcinoma cell line) cells, labeling Cdk4 with purified ab108357 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500), was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • Flow Cytometry (Intracellular) - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357)

    ab108357 staining CDK4in the human cell line MCF7 (human breast adenocarcinoma cell line) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabeled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357), expandable thumbnail

    Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357)

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357) at 1/10000 dilution

    Lane 1: HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg

    Lane 2: K562 (human chronic myelogenous leukemia cell line from bone marrow) cell lysate at 20 µg

    Lane 3: Ramos (human Burkitt's lymphoma cell line) cell lysate at 20 µg

    Secondary

    All lanes: Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size: 34 kDa

    Observed band size: 34 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357)

    ab108357 staining Cdk4 in wild-type HAP1 cells (top panel) and Cdk4 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), 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 ab108357 at 1/500 dilution and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 at 1/250 dilution (shown in pseudo colour red) 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

  • Flow Cytometry (Intracellular) - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357)

    Intracellular Flow Cytometry analysis of MCF7 (human breast adenocarcinoma cell line) cells labelling Cdk4 with purified ab108357 at 1/40 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357)

    Immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded human urothelial carcinoma tissue labelling Cdk4 with unpurified ab108357 at a dilution of 1/100.

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

  • Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357), expandable thumbnail
    This image is courtesy of a customer review submitted by Sonia Rocha

    Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357)

    All lanes: Western blot - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357) at 1/1000 dilution

    Lane 1: Human osteosarcoma whole cell lysate - control, non-targeting siRNA at 20 µg

    Lane 2: Human osteosarcoma whole cell lysate - siRNA for CDK4 at 20 µg

    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: 5s

  • OI-RD Scanning - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357), expandable thumbnail

    OI-RD Scanning - Anti-Cdk4 antibody [EPR4513-32-7] (ab108357)

    We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
    Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

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Product protocols

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