Overview

  • Product name
    Anti-Cyclin D1 antibody [SP4]
    See all Cyclin D1 primary antibodies
  • Description
    Rabbit monoclonal [SP4] to Cyclin D1
  • Tested applications
    Suitable for: ICC, ICC/IF, IHC-P, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide corresponding to Human Cyclin D1 (C terminal).

  • Epitope
    C-terminus
  • Positive control
    • Breast carcinomas, mantle cell lymphoma, MCF7 cell lysate IHC: Rat Esophagus (FFPE) ICC/IF: MCF7 cells, HAP1 cells (HAP1-CCND1 knockout cells used as negative cell line)

Properties

Applications

Our Abpromise guarantee covers the use of ab16663 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
ICC Use at an assay dependent concentration.
ICC/IF 1/250.
IHC-P 1/100.
WB 1/25 - 1/200. Detects a band of approximately 36 kDa (predicted molecular weight: 33 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Deparaffinization: Deparaffinize slides using xylene or xylene alternative and graded alcohols.

Antigen Retrieval: Boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min.

Primary Antibody Incubation: Incubate for 30 minutes at room temperature.

Slide Washing: Slides must be washed in between steps. Rinse slides with PBS/0.05% Tween.

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

Images



  • Predicted band size : 33 kDa

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: CCND1 (Cyclin D1) knockout HAP1 whole cell lysate (20 µg)
    Lane 3: A431 whole cell lysate (20 µg)

    Lanes 1 - 3: Merged signal (red and green). Green - ab16663 observed at 34 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab16663 was shown to specifically recognize CCND1 (Cyclin D1) in wild-type HAP1 cells as signal was lost at the expected MW in CCND1 (Cyclin D1) knockout cells. Wild-type and CCND1 (Cyclin D1) knockout samples were subjected to SDS-PAGE. Ab16663 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/200 dilution and 1/20,000 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 1/20,000 dilution for 1 hour at room temperature before imaging.

  • ab16663 staining Cyclin D1 in wild-type HAP1 cells (top panel) and CCND1 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), 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 ab16663 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudocolour 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.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • ab16663 staining Cyclin D1 in MCF7 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes 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 ab16663 at a working dilution of 1/250 and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-rabbit AlexaFluor® 488 (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • IHC image of ab16663 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 ab16663, 1:100 dilution, 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.

  • Immunohistochemical analysis of mouse testis tissue, staining Cyclin D1 with ab16663.

    Antigen retrieval was performed via Tris-EDTA buffer. Sections were blocked with 3% BSA and incubated with primary antibody (1/50) overnight at 4°C. An AlexaFluor®594-conjugated secondary antibody was used to detect staining.
  • Anti-Cyclin D1 antibody [SP4] (ab137875) at 1/5000 dilution + MCF-7 cell lysate

    Predicted band size : 33 kDa
  • Lane 1 : Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/200 dilution
    Lane 2 : Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/400 dilution

    Lane 1 : Whole cell lysate prepared from T24 bladder cancer cells
    Lane 2 : Whole cell lysate prepared from T24 bladder cancer cells

    Lysates/proteins at 25 µg per lane.

    Secondary
    Goat anti-rabbit IgG conjugated to HRP at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 33 kDa
    Observed band size : 33 kDa


    Exposure time : 10 minutes

    Image kindly supplied by Dr Karin Birkenkamp-Demtroeder through Abreview

    Gel run under denaturing conditions 4-12% gradient.

    See Abreview

  • Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/25 dilution + MCF7 cell lysate

    Predicted band size : 33 kDa
    Observed band size : 36 kDa (why is the actual band size different from the predicted?)
  • ab16663 staining Cyclin D1 in Human urinary tract tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% BSA for 30 minutes at room temperature; antigen retrieval was by heat mediation in citrate buffer. Samples were incubated with primary antibody (1/100 in PBS) for 1 hour. An undiluted HRP-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • Human mantle cell lymphoma stained with ab16663.

References

This product has been referenced in:
  • Tanaka T  et al. The efficacy of the cyclin-dependent kinase 4/6 inhibitor in endometrial cancer. PLoS One 12:e0177019 (2017). IHC-P ; Human . Read more (PubMed: 28472136) »
  • Ma L  et al. Long non-coding RNA XIST promotes cell growth and invasion through regulating miR-497/MACC1 axis in gastric cancer. Oncotarget 8:4125-4135 (2017). WB ; Human . Read more (PubMed: 27911852) »

See all 56 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (SW480 cells)
Loading amount
25 µg
Specification
SW480 cells
Gel Running Conditions
Reduced Denaturing (4-12%)
Username

Abcam user community

Verified customer

Submitted Jan 24 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (organ cultured corneal epithelium)
Specification
organ cultured corneal epithelium
Fixative
Formaldehyde
Permeabilization
Yes - 1% Triton
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 37°C
Username

Dr. Zhiguo He

Verified customer

Submitted Nov 06 2012

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (esophagus)
Specification
esophagus
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM citrate buffer
Permeabilization
No
Blocking step
DAKO serum free protein blocker as blocking agent for 20 minute(s) · Concentration: 0.25% · Temperature: 28°C
Username

Dr. J Chai

Verified customer

Submitted Apr 16 2012

Application
Western blot
Sample
Human Cell lysate - whole cell (Epithelial OVCAR cell line)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
100 µg
Treatment
Perifosine 0/12/24h
Specification
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 Oct 21 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Rat Placenta)
Antigen retrieval step
Enzymatic
Permeabilization
No
Specification
Rat Placenta
Blocking step
Serum as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 1.5%
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 26 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colon adenocarcinoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 0.01M citrate buffer
Permeabilization
No
Specification
Colon adenocarcinoma
Blocking step
(agent) for 20 minute(s) · Concentration: 100% · Temperature: 20°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 08 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (KMS12, U266, RPMI-8266, Daudi)
Permeabilization
Yes - 0.5% Triton X-100
Specification
KMS12, U266, RPMI-8266, Daudi
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Ms. Natalie Erdmann

Verified customer

Submitted Sep 10 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (Human stromal cell line (WPMY-1))
Gel Running Conditions
Reduced Denaturing (gel 10%)
Loading amount
50 µg
Treatment
various conditioned media
Specification
Human stromal cell line (WPMY-1)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Miss. Junghyun Kim

Verified customer

Submitted Jul 01 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Rat Cell (brain section)
Specification
brain section
Permeabilization
Yes - triton x-100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 17 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (fibrotic mouse liver)
Loading amount
20 µg
Specification
fibrotic mouse liver
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Nov 16 2012

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