Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP272Y] to Cyclin D1 - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free
    See all Cyclin D1 primary antibodies

  • Description

    Rabbit monoclonal [EP272Y] to Cyclin D1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: HeLa, HAP1, MCF-7, and A431 cell lysates. ICC/IF: MCF7 cells. IHC-P: Human kidney carcinoma and human hepatocellular carcinoma tissues.

  • General notes

    Ab227561 is the carrier-free version of ab40754. 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

    ab227561 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
    Western blot - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
    All lanes : Anti-Cyclin D1 antibody [EP272Y] (ab40754) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : CCND1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 33 kDa
    Observed band size: 36 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab40754).

      Lanes 1- 2: Merged signal (red and green). Green - ab40754 observed at 36 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.

     ab40754 was shown to react with Cyclin D1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255348 (knockout cell lysate ab263808) was used. Wild-type HeLa and CCND1 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. ab40754 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
    Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)

    ab40754 staining cyclin D1 in MCF7 cells treated with KN-93 (water soluble) (ab120980), by ICC/IF. Decrease in cyclin D1 expression correlates with increased concentration of KN-93 (water soluble), as described in literature.
    The cells were incubated at 37°C for 24 hours in media containing different concentrations of ab120980 (KN-93 (water soluble)) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab40754 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A goat anti-rabbit DyLight 488 goat anti-rabbit secondary antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40754).

  • Western blot - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
    Western blot - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
    All lanes : Anti-Cyclin D1 antibody [EP272Y] (ab40754) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : CCND1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 33 kDa
    Observed band size: 36 kDa why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab40754).

    Lanes 1- 2: Merged signal (red and green). Green - ab40754 observed at 36 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab40754 was shown to react with Cyclin D1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261760 (knockout cell lysate ab256864) was used. Wild-type HeLa and CCND1 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. ab40754 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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.

  • Western blot - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
    Western blot - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
    All lanes : Anti-Cyclin D1 antibody [EP272Y] (ab40754) at 1/1000 dilution

    Lane 2 : Wild-type HAP1 whole cell lysate
    Lane 3 : Cyclin D1 knockout HAP1 whole cell lysate
    Lane 4 : A431 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 33 kDa



    This WB data was generated using the same anti-Cyclin D1 antibody clone, EP272Y, in a different buffer formulation (cat# ab40754).

    Lanes 1 - 4: Merged signal (red and green). Green - ab40754 observed at 35 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab40754 was shown to recognize Cyclin D1 when HAP1 knockout samples were used, along with additional cross-reactive bands. Wild-type and HAP1 knockout samples were subjected to SDS-PAGE. ab40754 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)

    Immunohistochemical analysis of paraffin-embedded human kidney carcinoma using anti-Cyclin D1 RabMAb (ab40754).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 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 (ab40754).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)

    Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using anti-Cyclin D1 RabMAb (ab40754).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    This IHC data was generated using the same anti-Cyclin D1 antibody clone, EP272Y, in a different buffer formulation (cat# ab40754).

  • Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
    Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)

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