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Cell Biology Cell Cycle Cyclins Cyclin D Family
使用敲除细胞株进行验证重组RabMAb

重组Anti-Cyclin D1抗体[SP4] (ab16663)

  • Datasheet
  • SDS
Reviews (23)Q&A (5)References (358)

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Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Flow Cytometry (Intracellular) - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Flow Cytometry (Intracellular) - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Flow Cytometry (Intracellular) - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)
  • Anti-Cyclin D1 antibody [SP4] (ab16663)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [SP4] to Cyclin D1
  • Suitable for: ICC/IF, Flow Cyt (Intra), WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free

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概述

  • 产品名称

    Anti-Cyclin D1抗体[SP4]
    参阅全部 Cyclin D1 一抗
  • 描述

    兔单克隆抗体[SP4] to Cyclin D1
  • 宿主

    Rabbit
  • 经测试应用

    适用于: ICC/IF, Flow Cyt (Intra), WB, IHC-Pmore details
  • 种属反应性

    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide. This information is considered to be commercially sensitive.

  • 表位

    C-terminus
  • 阳性对照

    • WB: MCF7, Hap1, A431 and HeLa, Nuero-2a, NIH/3T3, C6 cell lysates. IHC (FFPE): Human normal tonsil; breast carcinoma; mantle cell lymphoma; rat esophagus. ICC/IF: MCF7 cells, C6, Neuro-2a and HAP1 cells (HAP1-CCND1 knockout cells used as negative cell line). Flow Cyt (intra): MCF7, NIH/3T3 and C6 cells.
  • 常规说明

    This product was switched from a hybridoma to recombinant production method on 22nd January 2019.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • 存储溶液

    pH: 7.20
    Preservative: 0.1% Sodium azide
    Constituents: 1% BSA, PBS
  • Concentration information loading...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    SP4
  • 同种型

    IgG
  • 研究领域

    • 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

相关产品

  • Alternative Versions

    • Anti-Cyclin D1 antibody [SP4] - BSA and Azide free (ab239794)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • 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)
  • Recombinant Protein

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

    • Recombinant Human Cyclin D1 protein (ab85247)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab16663于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC/IF (5)
1/50 - 1/250.
Flow Cyt (Intra)
1/30.
WB (12)
1/25 - 1/200. Detects a band of approximately 36 kDa (predicted molecular weight: 33 kDa).
IHC-P (5)
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.

说明
ICC/IF
1/50 - 1/250.
Flow Cyt (Intra)
1/30.
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.

靶标

  • 功能

    Essential for the control of the cell cycle at the G1/S (start) transition.
  • 疾病相关

    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.
  • 序列相似性

    Belongs to the cyclin family. Cyclin D subfamily.
  • 翻译后修饰

    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.
  • 细胞定位

    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
  • 数据库链接

    • 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
  • 别名

    • 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

图片

  • Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
    All lanes : Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/200 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?



    Lanes 1- 2: Merged signal (red and green). Green - ab16663 observed at 36 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.

     ab16663 was shown to react with CCND1 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. ab16663 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 200 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)

    IHC image of ab16663 staining Cyclin D1 in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. 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. The inset secondary-only control image is taken from an identical assay without primary antibody.

    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

    This image was generated from the hybridoma version.

     

  • Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/200 dilution + Wild-type HeLa cell lysate at 20 µg

    Performed under reducing conditions.

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



    Lanes 1- 2: Merged signal (red and green). Green - ab16663 observed at 36 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab16663 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. ab16663 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 200 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 [SP4] (ab16663)
    Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [SP4] (ab16663)

    Immunocytochemistry/ Immunofluorescence analysis of C6 (rat glial tumor glial cell) cells labeling Cyclin D1 with purified ab16663 at 1/50 (5.42µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    This image was generated from the hybridoma version.

     

  • Flow Cytometry (Intracellular) - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Flow Cytometry (Intracellular) - Anti-Cyclin D1 antibody [SP4] (ab16663)

    Intracellular flow cytometry analysis of MCF-7 (human breast carcinoma) labeling Cyclin D1 with purified ab16663 at 1/30 dilution (9.03 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlableled control - Unlabelled cells (blue). This image was generated from the hybridoma version.

  • Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
    All lanes : Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/1000 dilution

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma epithelial cell) cell lysate
    Lane 3 : CCND1 KO HAP1 cell lysate
    Lane 4 : HAP1 (Human chronic myelogenous leukemia near-haploid cell line) cell lysate
    Lane 5 : Neuro-2a (Mouse neuroblastoma neuroblast) cell lysate
    Lane 6 : NIH/3T3 (Mouse embryonic fibroblast) cell lysate
    Lane 7 : C6 (Rat glial tumor glial cell) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

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


    Exposure time: 2 seconds
  • Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
    All lanes : Anti-Cyclin D1 antibody [SP4] (ab16663)

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

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 33 kDa



    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.

    This image was generated from the hybridoma version.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)

    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.

    This image was generated from the hybridoma version.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [SP4] (ab16663)

    Immunocytochemistry/ Immunofluorescence analysis of Neuro-2a (mouse neuroblastoma neuroblast) cells labeling Cyclin D1µ with purified ab16663 at 1/50 (5.42µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    This image was generated from the hybridoma version.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [SP4] (ab16663)

    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).

    This image was generated from the hybridoma version.

     

  • Flow Cytometry (Intracellular) - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Flow Cytometry (Intracellular) - Anti-Cyclin D1 antibody [SP4] (ab16663)

    Intracellular flow cytometry analysis of C6 (rat glioma) labeling Cyclin D1 with purified ab16663 at 1/30 dilution (9.03 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlableled control - Unlabelled cells (blue). This image was generated from the hybridoma version.

  • Flow Cytometry (Intracellular) - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Flow Cytometry (Intracellular) - Anti-Cyclin D1 antibody [SP4] (ab16663)

    Intracellular flow cytometry analysis of NIH/3T3 (mouse embryo) labeling Cyclin D1 with purified ab16663 at 1/30 dilution (9.03 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlableled control - Unlabelled cells (blue).  This image was generated from the hybridoma version.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Immunocytochemistry/ Immunofluorescence - Anti-Cyclin D1 antibody [SP4] (ab16663)

    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).

    This image was generated from the hybridoma version.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)Image from McIver SC et al., PLoS One. 2012;7(4):e35553. Epub 2012 Apr 20. Fig 7.; doi:10.1371/journal.pone.0035553; April 20, 2012, PLoS ONE 7(4): e35553.

    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.

    This image was generated from the hybridoma version.

     

  • Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Anti-Cyclin D1 antibody [SP4] (ab137875) at 1/5000 dilution + MCF-7 cell lysate

    Predicted band size: 33 kDa



    This image was generated from the hybridoma version.

     

  • Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)Image kindly supplied by Dr Karin Birkenkamp-Demtroeder through Abreview
    Lane 1 : Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/200 dilution
    Lane 2 : Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/400 dilution

    All lanes : Whole cell lysate prepared from T24 bladder cancer cells

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : 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


    Gel run under denaturing conditions 4-12% gradient.

    This image was generated from the hybridoma version.

     

    See Abreview

  • Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Western blot - Anti-Cyclin D1 antibody [SP4] (ab16663)
    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?



    This image was generated from the hybridoma version.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)This image is courtesy of an Abreview submitted by Karin Birkenkamp-Demtroeder

    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.

    This image was generated from the hybridoma version.

     

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin D1 antibody [SP4] (ab16663)

    Human mantle cell lymphoma stained with ab16663.

    This image was generated from the hybridoma version.

     

  • Anti-Cyclin D1 antibody [SP4] (ab16663)
    Anti-Cyclin D1 antibody [SP4] (ab16663)

实验方案

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (358)

发表研究结果有使用 ab16663?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab16663 被引用在 358 文献中.

  • Di Leo L  et al. Loss of Ambra1 promotes melanoma growth and invasion. Nat Commun 12:2550 (2021). WB, IHC-P . PubMed: 33953176
  • Sun L & Zhang X Report of a rare case of congenital mitral valve prolapse with chronic kidney disease--reconsidered genotype-phenotypic correlations. Mol Genet Genomic Med 9:e1558 (2021). PubMed: 33225636
  • Ma Y  et al. SphK1 promotes development of non-small cell lung cancer through activation of STAT3. Int J Mol Med 47:374-386 (2021). PubMed: 33236138
  • Wang S  et al. lncRNA SNHG4 promotes cell proliferation, migration, invasion and the epithelial-mesenchymal transition process via sponging miR-204-5p in gastric cancer. Mol Med Rep 23:N/A (2021). PubMed: 33236157
  • Luo J  et al. miR-576-5p promotes epithelial-to-mesenchymal transition in colorectal cancer by targeting the Wnt5a-mediated Wnt/ß-catenin signaling pathway. Mol Med Rep 23:N/A (2021). PubMed: 33300054
View all Publications for this product

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1-10 of 28 Abreviews or Q&A

Western blot abreview for Anti-Cyclin D1 antibody [SP4]

Excellent
Abreviews
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abreview image
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%)
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Jan 24 2013

Immunocytochemistry/ Immunofluorescence abreview for Anti-Cyclin D1 antibody [SP4]

Good
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Abreviews
abreview image
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
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The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Zhiguo He

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提交于 Nov 06 2012

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cyclin D1 antibody [SP4]

Excellent
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Abreviews
abreview image
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
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The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. J Chai

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提交于 Apr 16 2012

Western blot abreview for Anti-Cyclin D1 antibody [SP4]

Excellent
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Abreviews
abreview image
Application
Western blot
Sample
Monkey Cell lysate - whole cell (COS-1 cells)
Gel Running Conditions
Reduced Denaturing (4-15%)
Loading amount
25 µg
Specification
COS-1 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Apr 08 2022

Western blot abreview for Anti-Cyclin D1 antibody [SP4]

Poor
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Abreviews
abreview image
Application
Western blot
Sample
Dog Cell lysate - whole cell (MDCK cells)
Gel Running Conditions
Reduced Denaturing (4-15%)
Loading amount
25 µg
Specification
MDCK cells
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Mar 04 2022

Western blot abreview for Anti-Cyclin D1 antibody [SP4]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rat Cell lysate - whole cell (3611-RF cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
3611-RF cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Mar 04 2022

Western blot abreview for Anti-Cyclin D1 antibody [SP4]

Excellent
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Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEFs)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
MEFs
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Mar 04 2022

Western blot abreview for Anti-Cyclin D1 antibody [SP4]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (MIA PaCa-2 cells)
Gel Running Conditions
Reduced Denaturing (4-15%)
Loading amount
25 µg
Specification
MIA PaCa-2 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Mar 04 2022

Western blot abreview for Anti-Cyclin D1 antibody [SP4]

Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse neural stem cell)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
40 µg
Specification
mouse neural stem cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 15°C
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The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Robert Kupp

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提交于 Sep 17 2019

Immunocytochemistry/ Immunofluorescence abreview for Anti-Cyclin D1 antibody [SP4]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (RPE1-hTERT cells)
Permeabilization
Yes - 0.2% Triton X-100
Specification
RPE1-hTERT cells
Blocking step
10% FBS, 1% BSA, 0.1% Triton X-100, 0.01% NaN3 in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Jan 19 2019

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