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  1. Link

    cyclin-b1-antibody-y106-ab32053.pdf

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Cell Biology Cell Cycle Cyclins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Cyclin B1 antibody [Y106] (ab32053)

  • Datasheet
  • SDS
Reviews (4)Q&A (1)References (129)

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Western blot - Anti-Cyclin B1 antibody [Y106] (ab32053)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [Y106] (ab32053)
  • Flow Cytometry (Intracellular) - Anti-Cyclin B1 antibody [Y106] (ab32053)
  • Western blot - Anti-Cyclin B1 antibody [Y106] (ab32053)
  • Immunoprecipitation - Anti-Cyclin B1 antibody [Y106] (ab32053)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin B1 antibody [Y106] (ab32053)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [Y106] (ab32053)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin B1 antibody [Y106] (ab32053)
  • Flow Cytometry (Intracellular) - Anti-Cyclin B1 antibody [Y106] (ab32053)
  • Western blot - Anti-Cyclin B1 antibody [Y106] (ab32053)
  • Anti-Cyclin B1 antibody [Y106] (ab32053)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y106] to Cyclin B1
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P, IP
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-Cyclin B1 antibody [Y106]
    See all Cyclin B1 primary antibodies
  • Description

    Rabbit monoclonal [Y106] to Cyclin B1
  • Host species

    Rabbit
  • Specificity

    This antibody is specific for Human cyclin B1. It does not cross-react with other cyclin family members.
  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Epitope

    ab32053 reacts with an epitope located in teh C terminal region of Cyclin B1.
  • Positive control

    • Jurkat and HeLa whole cell lysate (ab150035).
  • General notes

    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.

    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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y106
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Alternative Versions

    • Anti-Cyclin B1 antibody [Y106] - BSA and Azide free (ab156447)
    • HRP Anti-Cyclin B1 antibody [Y106] (ab193977)
    • PE Anti-Cyclin B1 antibody [Y106] (ab210615)
    • Alexa Fluor® 555 Anti-Cyclin B1 antibody [Y106] (ab214381)
  • 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)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab7899)
  • Recombinant Protein

    • Recombinant Human Cyclin B1 protein (ab128445)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32053 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
Flow Cyt (Intra)
1/400.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  For unpurified use at 1/20.

ICC/IF (1)
1/100.

For unpurified use at 1/50.

WB (2)
1/50000. Predicted molecular weight: 58 kDa.

For unpurified use at 1/3000 - 1/20000.

IHC-P (1)
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/20.

For unpurified use at 1/100.

Notes
Flow Cyt (Intra)
1/400.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  For unpurified use at 1/20.

ICC/IF
1/100.

For unpurified use at 1/50.

WB
1/50000. Predicted molecular weight: 58 kDa.

For unpurified use at 1/3000 - 1/20000.

IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/20.

For unpurified use at 1/100.

Target

  • Function

    Essential for the control of the cell cycle at the G2/M (mitosis) transition.
  • Sequence similarities

    Belongs to the cyclin family. Cyclin AB subfamily.
  • Developmental stage

    Accumulates steadily during G2 and is abruptly destroyed at mitosis.
  • Post-translational
    modifications

    Ubiquitinated by the SCF(NIPA) complex during interphase, leading to its destruction. Not ubiquitinated during G2/M phases.
  • Cellular localization

    Cytoplasm. Nucleus. Cytoplasm > cytoskeleton > centrosome.
  • Target information above from: UniProt accession P14635 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 891 Human
    • Omim: 123836 Human
    • SwissProt: P14635 Human
    • Unigene: 23960 Human
    • Alternative names

      • CCNB 1 antibody
      • CCNB antibody
      • ccnb1 antibody
      • CCNB1_HUMAN antibody
      • Cyclin B1 antibody
      • G2 mitotic specific cyclin B1 antibody
      • G2/mitotic-specific cyclin-B1 antibody
      see all

    Images

    • Western blot - Anti-Cyclin B1 antibody [Y106] (ab32053)
      Western blot - Anti-Cyclin B1 antibody [Y106] (ab32053)

      Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
      Lane 2: CCNB1 knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lanes 1 - 3: Merged signal (red and green). Green - ab32053 observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.

      Unpurified ab32053 was shown to specifically react with CCNB1 in wild-type HAP1 cells. No band was observed when CCNB1 knockout samples were examined. Wild-type and CCNB1 knockout samples were subjected to SDS-PAGE. Ab32053 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/3000 dilution and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [Y106] (ab32053)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [Y106] (ab32053)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cervical carcinoma tissue sections labeling Cyclin B1 with Purified ab32053 at 1:250 dilution (1.47 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

    • Flow Cytometry (Intracellular) - Anti-Cyclin B1 antibody [Y106] (ab32053)
      Flow Cytometry (Intracellular) - Anti-Cyclin B1 antibody [Y106] (ab32053)

      Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Cyclin B1 with purified ab32053 at 1/400 dilution (1µg/ml) (Right). Cells were fixed with 80% Methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Left). Cells were pre-treated with 20µg/ml RNaseA for 30min to minimize the binding between PI and RNA.Then intracellular stained with ab32053 and PI. 

       

       

    • Western blot - Anti-Cyclin B1 antibody [Y106] (ab32053)
      Western blot - Anti-Cyclin B1 antibody [Y106] (ab32053)
      All lanes : Anti-Cyclin B1 antibody [Y106] (ab32053) at 1/50000 dilution (purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 58 kDa
      Observed band size: 58 kDa



      Blocking and diluting buffer: 5% NFDM/TBST

    • Immunoprecipitation - Anti-Cyclin B1 antibody [Y106] (ab32053)
      Immunoprecipitation - Anti-Cyclin B1 antibody [Y106] (ab32053)

      ab32053 (purified) at 1:20 dilution (2μg) immunoprecipitating Cyclin B1 in Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate.

      Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10μg
      Lane 2 (+): ab32053 & Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32053 in Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate

      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin B1 antibody [Y106] (ab32053)
      Immunocytochemistry/ Immunofluorescence - Anti-Cyclin B1 antibody [Y106] (ab32053)

      Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Cyclin B1 with Purified ab32053 at 1:100 dilution. Cells were fixed in 100% Methanol. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [Y106] (ab32053)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [Y106] (ab32053)

      Unpurified ab32053 at a 1:100 dilution staining Human cyclin B1 in human skin carcinoma, using Immunohistochemistry, Paraffin Embedded Tissue.

    • Immunocytochemistry/ Immunofluorescence - Anti-Cyclin B1 antibody [Y106] (ab32053)
      Immunocytochemistry/ Immunofluorescence - Anti-Cyclin B1 antibody [Y106] (ab32053)This image is courtesy of an Abreview submitted by Stephanie Hilss

      Unpurified ab32053 staining Cyclin B1 in the U2OS cell line from human by ICC/IF (Immunocytochemistry/immunofluorescence).Cells were fixed with formaldehyde,permeabilized with 1% Triton X-100 in PBS and blocked with 1% BSA for 1 hour at 37°C.Alexa Fluor® 594-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

      See Abreview

    • Flow Cytometry (Intracellular) - Anti-Cyclin B1 antibody [Y106] (ab32053)
      Flow Cytometry (Intracellular) - Anti-Cyclin B1 antibody [Y106] (ab32053)

      Overlay histogram showing Jurkat cells stained with unpurified ab32053 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32053, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. 

       

       

    • Western blot - Anti-Cyclin B1 antibody [Y106] (ab32053)
      Western blot - Anti-Cyclin B1 antibody [Y106] (ab32053)
      All lanes : Anti-Cyclin B1 antibody [Y106] (ab32053) at 1/20000 dilution (unpurified)

      Lane 1 : Hela cell lysate.
      Lane 2 : Jurkat cell lysate.

      Predicted band size: 58 kDa

    • Anti-Cyclin B1 antibody [Y106] (ab32053)
      Anti-Cyclin B1 antibody [Y106] (ab32053)

    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

    • SDS download

    • Datasheet download

      Download

    References (129)

    Publishing research using ab32053? Please let us know so that we can cite the reference in this datasheet.

    ab32053 has been referenced in 129 publications.

    • Zhang Y  et al. Long Non-coding RNA CASC15 Promotes Intrahepatic Cholangiocarcinoma Possibly through Inducing PRDX2/PI3K/AKT Axis. Cancer Res Treat 53:184-198 (2021). PubMed: 33017884
    • Velásquez ZD  et al. Eimeria bovis infections induce G1 cell cycle arrest and a senescence-like phenotype in endothelial host cells. Parasitology 148:341-353 (2021). PubMed: 33100232
    • Wu F  et al. Bleomycin A5 suppresses Drp1-mediated mitochondrial fission and induces apoptosis in human nasal polyp-derived fibroblasts. Int J Mol Med 47:346-360 (2021). PubMed: 33236140
    • Yang T  et al. Punicalin Alleviates OGD/R-Triggered Cell Injury via TGF-ß-Mediated Oxidative Stress and Cell Cycle in Neuroblastoma Cells SH-SY5Y. Evid Based Complement Alternat Med 2021:6671282 (2021). PubMed: 33628309
    • Kim CH  et al. Bisphenol A Exposure Changes the Transcriptomic and Proteomic Dynamics of Human Retinoblastoma Y79 Cells. Genes (Basel) 12:N/A (2021). PubMed: 33670352
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review

    Filter by Application

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    1-4 of 4 Abreviews

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Cyclin B1 antibody [Y106]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (U2OS)
    Permeabilization
    Yes - 0.1% TritonX-100
    Specification
    U2OS
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More

    DR. Stephanie Hills

    Verified customer

    Submitted Apr 27 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cyclin B1 antibody [Y106]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    H2O2 as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA, pH 8.6, 20 min 100C
    Sample
    Human Tissue sections (tonsil)
    Specification
    tonsil
    Permeabilization
    No
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 06 2014

    Western blot abreview for Anti-Cyclin B1 antibody [Y106]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Sample
    Human Cell lysate - whole cell (MCF7 cells)
    Specification
    MCF7 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 11 2013

    Western blot abreview for Anti-Cyclin B1 antibody [Y106]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa cells)
    Loading amount
    30 µg
    Specification
    HeLa cells
    Treatment
    double thymidine synchronization
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C
    Read More

    Abcam user community

    Verified customer

    Submitted May 14 2009

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