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  1. Link

    cyclin-d1-antibody-ep272y-bsa-and-azide-free-ab227561.pdf

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Cell Biology Cell Cycle Cyclins Cyclin D Family
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
  • Western blot - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
  • Western blot - 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)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561)
  • 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

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Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
Protein
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Recombinant Human Cyclin D1 protein (ab85247)

View more associated products

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

    Synthetic peptide corresponding to residues near the N-terminus (Human)

  • 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 <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.20
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP272Y
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cyclins
    • Cyclin D Family
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Cyclins
    • Cyclin D Family
    • Cancer
    • Cell cycle
    • Cyclins
    • Cyclin D family

Associated products

  • Alternative Versions

    • Anti-Cyclin D1 antibody [EP272Y] (ab40754)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
  • Conjugation kits

    • Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CCND1 (Cyclin D1) knockout HeLa cell line (ab255348)
    • Human CCND1 (Cyclin D1) knockout HeLa cell line (ab261760)
  • KO cell lysates

    • Human CCND1 (Cyclin D1) knockout HeLa cell lysate (ab256864)
    • Human CCND1 (Cyclin D1) knockout HeLa cell lysate (ab263808)
  • Positive Controls

    • HeLa membrane extract lysate (ab29547)
  • Recombinant Protein

    • Recombinant Human Cyclin D1 protein (ab85247)
  • Related Products

    • KN-93 (water soluble), CaMK II inhibitor (ab120980)

Applications

Our Abpromise guarantee covers the use of ab227561 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 Use at an assay dependent concentration. Detects a band of approximately 35 kDa (predicted molecular weight: 33 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    • Function

      Essential for the control of the cell cycle at the G1/S (start) transition.
    • Involvement in disease

      Note=A chromosomal aberration involving CCND1 may be a cause of B-lymphocytic malignancy, particularly mantle-cell lymphoma (MCL). Translocation t(11;14)(q13;q32) with immunoglobulin gene regions. Activation of CCND1 may be oncogenic by directly altering progression through the cell cycle.
      Note=A chromosomal aberration involving CCND1 may be a cause of parathyroid adenomas. Translocation t(11;11)(q13;p15) with the parathyroid hormone (PTH) enhancer.
      Defects in CCND1 are a cause of multiple myeloma (MM) [MIM:254500]. MM is a malignant tumor of plasma cells usually arising in the bone marrow and characterized by diffuse involvement of the skeletal system, hyperglobulinemia, Bence-Jones proteinuria and anemia. Complications of multiple myeloma are bone pain, hypercalcemia, renal failure and spinal cord compression. The aberrant antibodies that are produced lead to impaired humoral immunity and patients have a high prevalence of infection. Amyloidosis may develop in some patients. Multiple myeloma is part of a spectrum of diseases ranging from monoclonal gammopathy of unknown significance (MGUS) to plasma cell leukemia. Note=A chromosomal aberration involving CCND1 is found in multiple myeloma. Translocation t(11;14)(q13;q32) with the IgH locus.
    • Sequence similarities

      Belongs to the cyclin family. Cyclin D subfamily.
    • Post-translational
      modifications

      Phosphorylation at Thr-286 by MAP kinases is required for ubiquitination and degradation following DNA damage. It probably plays an essential role for recognition by the FBXO31 component of SCF (SKP1-cullin-F-box) protein ligase complex.
      Ubiquitinated, primarily as 'Lys-48'-linked polyubiquitination. Ubiquitinated by a SCF (SKP1-CUL1-F-box protein) ubiquitin-protein ligase complex containing FBXO4 and CRYAB (By similarity). Following DNA damage it is ubiquitinated by some SCF (SKP1-cullin-F-box) protein ligase complex containing FBXO31. Ubiquitination leads to its degradation and G1 arrest. Deubiquitinated by USP2; leading to stabilize it.
    • Cellular localization

      Nucleus.
    • Target information above from: UniProt accession P24385 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 595 Human
      • Entrez Gene: 12443 Mouse
      • Entrez Gene: 58919 Rat
      • Omim: 168461 Human
      • SwissProt: P24385 Human
      • SwissProt: P25322 Mouse
      • SwissProt: P39948 Rat
      • Unigene: 523852 Human
      • Unigene: 667996 Human
      • Unigene: 273049 Mouse
      • Unigene: 22279 Rat
      see all
    • Alternative names

      • AI327039 antibody
      • B cell CLL/lymphoma 1 antibody
      • B cell leukemia 1 antibody
      • B cell lymphoma 1 protein antibody
      • B-cell lymphoma 1 protein antibody
      • BCL 1 antibody
      • BCL-1 antibody
      • BCL-1 oncogene antibody
      • BCL1 antibody
      • BCL1 oncogene antibody
      • ccnd1 antibody
      • CCND1/FSTL3 fusion gene antibody
      • CCND1/FSTL3 fusion gene, included antibody
      • CCND1/IGHG1 fusion gene antibody
      • CCND1/IGHG1 fusion gene, included antibody
      • CCND1/IGLC1 fusion gene antibody
      • CCND1/IGLC1 fusion gene, included antibody
      • CCND1/PTH fusion gene antibody
      • CCND1/PTH fusion gene, included antibody
      • CCND1_HUMAN antibody
      • cD1 antibody
      • Cyl 1 antibody
      • D11S287E antibody
      • G1/S specific cyclin D1 antibody
      • G1/S-specific cyclin-D1 antibody
      • Parathyroid adenomatosis 1 antibody
      • PRAD1 antibody
      • PRAD1 oncogene antibody
      • U21B31 antibody
      see all

    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)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (8)

    Publishing research using ab227561? Please let us know so that we can cite the reference in this datasheet.

    ab227561 has been referenced in 8 publications.

    • Fischer MM  et al. WNT antagonists exhibit unique combinatorial antitumor activity with taxanes by potentiating mitotic cell death. Sci Adv 3:e1700090 (2017). PubMed: 28691093
    • Wu Y  et al. Low expression of secreted frizzled-related protein 2 and nuclear accumulation of ß-catenin in aggressive nonfunctioning pituitary adenoma. Oncol Lett 12:199-206 (2016). WB ; Human . PubMed: 27347125
    • Wang T  et al. Inhibition of transient receptor potential channel 5 reverses 5-Fluorouracil resistance in human colorectal cancer cells. J Biol Chem 290:448-56 (2015). PubMed: 25404731
    • He X  et al. Functional repair of p53 mutation in colorectal cancer cells using trans-splicing. Oncotarget 6:2034-45 (2015). Human . PubMed: 25576916
    • Dorfman T  et al. Enhanced intestinal epithelial cell proliferation in diabetic rats correlates with ß-catenin accumulation. J Endocrinol 226:135-43 (2015). PubMed: 26297291
    • Wu J  et al. MicroRNA-188 suppresses G1/S transition by targeting multiple cyclin/CDK complexes. Cell Commun Signal 12:66 (2014). WB ; Human . PubMed: 25304455
    • Gui J  et al. Enormous influence of TNIK knockdown on intracellular signals and cell survival. Hum Cell 24:121-6 (2011). PubMed: 21710359
    • Quiles I  et al. Mutational analysis of progesterone receptor functional domains in stable cell lines delineates sets of genes regulated by different mechanisms. Mol Endocrinol 23:809-26 (2009). WB ; Human . PubMed: 19299443

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