Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab134175 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 1/10000 - 1/50000. Predicted molecular weight: 34 kDa.
IP 1/30.
ICC/IF 1/50.
IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Target

  • FunctionEssential for the control of the cell cycle at the G1/S (start) transition.
  • Involvement in diseaseNote=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 similaritiesBelongs 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 localizationNucleus.
  • Information by UniProt
  • Database links
  • 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, included antibody
    • CCND1/IGHG1 fusion gene, included antibody
    • CCND1/IGLC1 fusion gene, included 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

Anti-Cyclin D1 antibody [EPR2241] images

  • Unpurified ab134175 staining Cyclin D1 in MCF7 cells treated with KN-93 (ab120980). The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab134175 at 10μg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an Goat anti-Rabbit Alexa 488 secondary (ab150081) at 2 μg/ml (shown in green) and Goat anti-Mouse Alexa 594 secondary (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

    Negative controls: 1– Rabbit primary and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

  • Immunohistochemical analysis of paraffin-embedded Human mantle cell lymphoma tissue labelling Cyclin D1 with unpurified ab134175 at 1/100 dilution.

  • IHC image of ab134175 staining Cyclin D1 in human esophagus formalin fixed paraffin embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab134175, 5μg/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • IHC image of ab134175 staining Cyclin D1 in rat esophagus formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab134175, 5μg/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-Cyclin D1 antibody [EPR2241] (ab134175) at 1/10000 dilution (purified)

    Lane 1 : Mouse kidney
    Lane 2 : Mouse spleen
    Lane 3 : Rat heart

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded human endometrial adenocarcinoma with purified ab134175 at a dilution of 1/100. A HRP goat anti-rabbit (ab97051) was used as the secondary antibody at a dilution of 1/500 and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunofluorescent staining of Ramos cells (fixed in 4% PFA, permeabilized with 0.1% Triton X 100) using purified ab134175 at a dilution of 1/50. An Alexa Fluor® 488 goat anti-rabbit antibody was used as the secondary at a dilution of 1/500 and the cells were counter stained with DAPI. The negative control is shown in the bottom right hand panel - for the negative control, Alex Fluor® 594 goat anti-mouse was used at a dilution of 1/500.

  • All lanes : Anti-Cyclin D1 antibody [EPR2241] (ab134175) at 1/100000 dilution (purified)

    Lane 1 : MCF-7 cell lysate
    Lane 2 : LnCap cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • All lanes : Anti-Cyclin D1 antibody [EPR2241] (ab134175) at 1/20000 dilution (purified)

    Lane 1 : HeLa cell lysate
    Lane 2 : A431 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 34 kDa


    Exposure time : 3 minutes

    MG-63 cells were incubated at 37°C for 24h with vehicle control (0 μM) and different concentrations of 3-aminobenzamide (3-AB) (ab141069). Decreased expression of cyclin D1 (unpurified ab134175) in MG-63 cells correlates with an increase in 3-aminobenzamide (3-AB) concentration, as described in literature.


    Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10 μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with unpurified ab134175 at 1/500 dilution and ab8226 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit HRP secondary antibody (ab97051) at 1/10000 dilution and visualised using ECL development solution.

  • ab134175 (purified) at 1/30 immunoprecipitating cyclin D1 in A431 cells (Lane 1). For western blotting, a HRP-conjugated goat anti-rabbit IgG, was used as the secondary antibody (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Anti-Cyclin D1 antibody [EPR2241] (ab134175) at 1/10000 dilution (unpurified) + MCF7 cell lysate at 10 µg

    Secondary
    HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size : 34 kDa

    Secondary goat anti-rabbit HRP (ab6721)

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References for Anti-Cyclin D1 antibody [EPR2241] (ab134175)

This product has been referenced in:
  • Misiewicz-Krzeminska I  et al. Post-transcriptional Modifications Contribute to the Upregulation of Cyclin D2 in Multiple Myeloma. Clin Cancer Res 22:207-17 (2016). Read more (PubMed: 26341922) »
  • Asher M  et al. Ataxin-1 regulates proliferation of hippocampal neural precursors. Neuroscience N/A:N/A (2016). Read more (PubMed: 26876606) »

See all 44 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Permeabilization Yes - 0.5% Triton X100 in PBS
Fixative Paraformaldehyde
Username

Dr. Kirk McManus

Verified customer

Submitted Jan 22 2014

Application Western blot
Loading amount 15 µg
Gel Running Conditions Reduced Denaturing (13%)
Sample Human Cell lysate - whole cell (HeLa and MEF cell lysates)
Specification HeLa and MEF cell lysates
Treatment See ab136811
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Christian Marx

Verified customer

Submitted Dec 04 2013

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Human Cell lysate - whole cell (osteosarcoma cells)
Specification osteosarcoma cells
Treatment lane 1 control, lane 2 siRNA for Cyclin D1, 48h
Blocking step Milk as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 21°C
Username

Dr. Sonia Rocha

Verified customer

Submitted Sep 26 2014

Application Western blot
Loading amount 15 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Tissue lysate - whole (Heart)
Specification Heart
Blocking step superblock as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 25 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Sample Human Cell (MCF-7)
Specification MCF-7
Permeabilization Yes - 0.1% Triton X-100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 04 2014

Application Western blot
Loading amount 50000 cells
Gel Running Conditions Reduced Denaturing (10%)
Sample Human Cell lysate - whole cell (MCF-7, MDA-MB-231)
Specification MCF-7, MDA-MB-231
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Aug 04 2014

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"