Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4513-32-7] to Cdk4 - BSA and Azide free
  • Suitable for: WB, ICC/IF, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free
    See all Cdk4 primary antibodies

  • Description

    Rabbit monoclonal [EPR4513-32-7] to Cdk4 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HAP1 and HeLa cell lysates, MCF7 membrane extract lysate (ab29539). Flow Cyt: MCF-7 IHC-P: Human cervix carcinoma and human urothelial carcinoma. ICC/IF: MCF-7 and HAP1.

  • General notes

    ab213216 is the carrier-free version of ab108357 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    Ab213216 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information. Alternative versions available: Anti-Cdk4 antibody [EPR4513-32-7] (ab108357) - Knockout validated

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
    Western blot - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
    All lanes : Anti-Cdk4 antibody [EPR4513-32-7] (ab108357) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : CDK4 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

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



    This data was developed using the same antibody clone in a different buffer formulation (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 ab255378 (knockout 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 (ab216773) and 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.

  • Flow Cytometry - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
    Flow Cytometry - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)

    ab108357 staining CDK4 in the human cell line MCF-7(human breast carcinoma) by 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)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108357).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)

    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). 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108357).

  • Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
    Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)

    Immunocytochemical analysis of MCF7 cells, labeling Cdk4 with purified ab108357 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. 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. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108357).

  • Western blot - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
    Western blot - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
    Lanes 1-2 : Anti-Cdk4 antibody [EPR4513-32-7] (ab108357) at 1/1000 dilution
    Lanes 3-4 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution


    Lanes 1 & 3 & 5 : Wild-type HAP1 cell lysate
    Lanes 2 & 4 & 6 : CDK4 knockout HAP1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 34 kDa



    This data was developed using the same antibody clone in a different buffer formulation (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 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 hour at room temperature before imaging.

  • Flow Cytometry - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
    Flow Cytometry - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)

    Flow Cytometry analysis of MCF7 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108357).

  • Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
    Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)

    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 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) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108357).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108357).

  • OI-RD Scanning - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
    OI-RD Scanning - Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108357).

  • Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
    Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)

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