Recombinant
RabMAb

Anti-Cyclin D1 antibody [EPR2241] (ab134175)

Overview

  • Product name
    Anti-Cyclin D1 antibody [EPR2241]
    See all Cyclin D1 primary antibodies
  • Description
    Rabbit monoclonal [EPR2241] to Cyclin D1
  • Specificity
    The immunogen used for this product shares 66% homology with CCND2 (seven amino acid stretch with 100% homology). The cross-reactivity with this protein has not been confirmed experimentally.
  • Tested applications
    Suitable for: WB, IP, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Cyclin D1 (C terminal).
    Database link: P24385
    (Peptide available as ab188123)

  • Positive control
    • Human mantle cell lymphoma tissue; MCF7 cell lysate. ICC/IF: MCF7 cells treated with KN-93 IHC/P: Human esophagus (FFPE), Rat esophagus (FFPE)
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels.

    If you have any questions regarding this update, please contact our Scientific Support team.

    A trial size is available to purchase for this antibody.

     

    Alternative versions available:

    Anti-Cyclin D1 antibody (Alexa Fluor® 488) [EPR2241] (ab190194)
    Anti-Cyclin D1 antibody (Alexa Fluor® 647) [EPR2241] (ab190563)
    Anti-Cyclin D1 antibody (HRP) [EPR2241] (ab190564)

    Anti-Cyclin D1 antibody (Alexa Fluor® 594) [EPR2241] (ab203446)
    Anti-Cyclin D1 antibody (Alexa Fluor® 555) [EPR2241] (ab203448)
    Anti-Cyclin D1 antibody (Alexa Fluor® 568) [EPR2241] (ab203449)

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

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

  • 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.
  • 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

  • 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 counterstained 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.

  • 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.

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

  • 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.

  • 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

  • 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 hematoxylin 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

  • Immunocytochemical analysis of HeLa cells, labeling Cyclin D1 with ab134175 at a dilution of 1/200. Cells were paraformaldehyde fixed and permeabilized with 0.5% Triton X-100 in PBS. Incubation with the primary antibody was for 1 hour at 22°C. Cells were counterstained with DAPI following immunostaining.

    See Abreview

  • 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 hematoxylin 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.


  • 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.

  • 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)

  • Immunocytochemical analysis of human MCF7 cells, labeling Cyclin D1 with ab134175 at a dilution of 1/100. Cells were paraformaldehyde fixed and permeabilized with 0.1% Triton X-100. To avoid non-specific binding, cells were blocked with 1% Goat serum + 1% BSA in PBS for 1 hour at 22°C. Incubation with the primary antibody was for 16 hours at 4°C. An Alexa Fluor® 488 conjugated goat anti-rabbit IgG polyclonal antibody, diluted 1/500.

    See Abreview

  • 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:
  • Wu VC  et al. The prevalence of CTNNB1 mutations in primary aldosteronism and consequences for clinical outcomes. Sci Rep 7:39121 (2017). WB ; Human . Read more (PubMed: 28102204) »
  • Lu T  et al. Adamts18 deficiency promotes colon carcinogenesis by enhancing ß-catenin and p38MAPK/ERK1/2 signaling in the mouse model of AOM/DSS-induced colitis-associated colorectal cancer. Oncotarget 8:18979-18990 (2017). IHC ; Mouse . Read more (PubMed: 28145888) »

See all 71 Publications for this product

Product Wall

Filter by Application

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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
Sample
Human Cell lysate - whole cell (Human Epithelial OVCAR cell line)
Gel Running Conditions
Reduced Denaturing (10 % SDS-PAGE)
Loading amount
100 µg
Treatment
Perifosine 0-24 hr
Specification
Human Epithelial OVCAR cell line
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Apr 20 2017

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

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