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Cell Biology Cell Cycle Cyclins
使用敲除细胞株进行验证

Anti-Cyclin B1抗体[V152] (ab72)

  • Datasheet
Reviews (9)Q&A (1)References (79)

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Western blot - Anti-Cyclin B1 antibody [V152] (ab72)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [V152] (ab72)
  • Western blot - Anti-Cyclin B1 antibody [V152] (ab72)
  • Western blot - Anti-Cyclin B1 antibody [V152] (ab72)

Key features and details

  • Mouse monoclonal [V152] to Cyclin B1
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

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概述

  • 产品名称

    Anti-Cyclin B1抗体[V152]
    参阅全部 Cyclin B1 一抗
  • 描述

    小鼠单克隆抗体[V152] to Cyclin B1
  • 宿主

    Mouse
  • 经测试应用

    适用于: WB, IHC-Pmore details
    不适用于: IP
  • 种属反应性

    与反应: Human
    预测可用于: Mouse
  • 免疫原

    Fusion protein within Hamster Cyclin B1. The exact sequence is proprietary.
    Database link: P14635

  • 阳性对照

    • WB: HeLa, Daudi, K562, Jurkat and HEK-293 whole cell lysate. IHC-P: Human tonsil tissue. Flow Cytometry: HeLa cells.
  • 常规说明

    This antibody clone is manufactured by Abcam.

    This monoclonal antibody to cyclin B1 has been knockout validated in Western blot. The expected band for cyclin B1 was observed in wild type cells and the band was not seen in knockout cells.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • 纯度

    Protein G purified
  • 克隆

    单克隆
  • 克隆编号

    V152
  • 骨髓瘤

    Sp2
  • 同种型

    IgG1
  • 轻链类型

    unknown
  • 研究领域

    • 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

相关产品

  • Alternative Versions

    • Anti-Cyclin B1 antibody [V152] - BSA and Azide free (ab186926)
    • FITC Anti-Cyclin B1 antibody [V152] (ab187806)
    • HRP Anti-Cyclin B1 antibody [V152] (ab201853)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772)
    • Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
  • Recombinant Protein

    • Recombinant Human Cyclin B1 protein (ab128445)

应用

Our Abpromise guarantee covers the use of ab72 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 48 kDa).

Abcam recommends using 5% Milk as the blocking agent.

IHC-P Use a concentration of 5 µg/ml.
  • 应用说明
    Is unsuitable for IP.
  • 靶标

    • 功能

      Essential for the control of the cell cycle at the G2/M (mitosis) transition.
    • 序列相似性

      Belongs to the cyclin family. Cyclin AB subfamily.
    • 发展阶段

      Accumulates steadily during G2 and is abruptly destroyed at mitosis.
    • 翻译后修饰

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

      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
    • 数据库链接

      • Entrez Gene: 891 Human
      • Entrez Gene: 268697 Mouse
      • Omim: 123836 Human
      • SwissProt: P14635 Human
      • SwissProt: P24860 Mouse
      • Unigene: 23960 Human
      • Unigene: 260114 Mouse
      • Unigene: 380027 Mouse
      • Unigene: 482545 Mouse
      see all
    • 别名

      • 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

    图片

    • Western blot - Anti-Cyclin B1 antibody [V152] (ab72)
      Western blot - Anti-Cyclin B1 antibody [V152] (ab72)

      Lanes:
      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: Cyclin B1 knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: HEK293 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab72 observed at 55 kDa. Red - loading control, ab181602, observed at 37 kDa.

      ab72 detected the expected band for Cyclin B1 in wild type HAP1 cells and the band was not seen in Cyclin B1 knockout HAP1 cells. Wild-type and Cyclin B1 knockout samples were subjected to SDS-PAGE. Ab72 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [V152] (ab72)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [V152] (ab72)

      ab72 staining Cyclin B1 in human tonsil tissue - formalin-fixed paraffin-embedded section, 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 (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab72, 5 µg/ml, for 15 minutes 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.

      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.

    • Western blot - Anti-Cyclin B1 antibody [V152] (ab72)
      Western blot - Anti-Cyclin B1 antibody [V152] (ab72)
      All lanes : Anti-Cyclin B1 antibody [V152] (ab72) at 1 µg/ml

      Lane 1 : HeLa (Human epithelial carcinoma cell line) whole cell lysate
      Lane 2 : Daudi whole cell lysate (ab3951)
      Lane 3 : K562 (Human erythromyeloblastoid leukemia cell line) whole cell lysate
      Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) whole cell lysate
      Lane 5 : HEK-293 (Human embryonic kidney cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 48 kDa
      Observed band size: 55 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 36 kDa, 77 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 20 minutes


      Abcam recommends using milk as the blocking agent.
    • Western blot - Anti-Cyclin B1 antibody [V152] (ab72)
      Western blot - Anti-Cyclin B1 antibody [V152] (ab72)

      Lanes 1-9 shows the Cyclin B1 wave as HeLa cells enter and exit mitosis. HeLa cells were synchronised in the early S phase by double thymidine block then released to synchronously enter mitosis. Time points were taken at the points indicated.

      HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate loaded at 25 µg per lane. ab72 used at 1/500 in reducing conditions in conjunction with goat anti mouse (HRP) 1/20,000.

      This image is courtesy of an Abreview submitted on 12 September 2005. We do not have any further information relating to this image.

       

      See Abreview

    实验方案

    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    数据表及文件

    • Datasheet
  • 文献 (79)

    发表研究结果有使用 ab72?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab72 被引用在 79 文献中.

    • Luong XG  et al. Genome-wide analysis reveals a switch in the translational program upon oocyte meiotic resumption. Nucleic Acids Res 48:3257-3276 (2020). PubMed: 31970406
    • Yao J  et al. circGFRA1 Enhances NSCLC Progression by Sponging miR-188-3p. Onco Targets Ther 13:549-558 (2020). PubMed: 32021297
    • Levasseur MD  et al. Aneuploidy in Oocytes Is Prevented by Sustained CDK1 Activity through Degron Masking in Cyclin B1. Dev Cell 48:672-684.e5 (2019). PubMed: 30745144
    • Daldello EM  et al. Cyclin B2 is required for progression through meiosis in mouse oocytes. Development 146:N/A (2019). PubMed: 30952665
    • Rodríguez C  et al. A novel human Cdh1 mutation impairs anaphase promoting complex/cyclosome activity resulting in microcephaly, psychomotor retardation, and epilepsy. J Neurochem 151:103-115 (2019). PubMed: 31318984
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-10 of 10 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cyclin B1 antibody [V152]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (human skin carcinoma)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 6.0 citric acid
    Permeabilization
    Yes - tween-20
    Specification
    human skin carcinoma
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Jun 22 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Cyclin B1 antibody [V152]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - tween-20
    Specification
    HeLa
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Jun 22 2015

    Western blot abreview for Anti-Cyclin B1 antibody [V152]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Jurkat cells)
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Loading amount
    30 µg
    Specification
    Jurkat 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

    提交于 Apr 22 2015

    Western blot abreview for Anti-Cyclin B1 antibody [V152]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    21 µg
    Gel Running Conditions
    Reduced Denaturing (10%)
    Sample
    Mouse Cell lysate - whole cell (MEL, K562)
    Specification
    MEL, K562
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    提交于 Nov 01 2013

    Flow Cytometry abreview for Anti-Cyclin B1 antibody [V152]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Preparation
    Cell harvesting/tissue preparation method: Trypsin/EDTA
    Sample buffer: Complete Media then PBS
    Fixation
    Formaldehyde
    Permeabilization
    Yes - 90% Methanol
    Read More

    Dr. Brandon White

    Verified customer

    提交于 Aug 29 2012

    Western blot abreview for Anti-Cyclin B1 antibody [V152]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Loading amount
    20 µg
    Specification
    HeLa
    Gel Running Conditions
    Reduced Denaturing (10%)
    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

    提交于 Aug 17 2011

    Western blot abreview for Anti-Cyclin B1 antibody [V152]

    Good
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Oocyte)
    Loading amount
    20 cells
    Specification
    Oocyte
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (10% Bis Tris Gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    提交于 Nov 18 2008

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Cyclin B1 antibody [V152]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Methanol
    Permeabilization
    No
    Read More

    Dr. Kirk Mcmanus

    Verified customer

    提交于 May 12 2008

    Western blot abreview for Anti-Cyclin B1 antibody [V152]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Specification
    HeLa
    Blocking step
    Serum as blocking agent for 45 minute(s) · Concentration: 5%
    Read More

    Abcam user community

    Verified customer

    提交于 Sep 12 2005

    Question

    I recently purchased the ab72 cyclin B1 antibody to be used in experiments as a mitotic marker. The experiments i was perfoming were time courses and drug arrests using hela cells. In blots this antibody seemed to detect two bands, one at the expected 62kDa and one above this at about 80. not only this but comparisons using S phase cells versus M phase cells showed no significant difference for cyclin B1, as a control phosphorylated histone H3 was used and was only detectable in M phase lysates as expected. From this data it would sugesst that this antibody does not recognise cyclin B1. Can you suggest any reasons for these observations.

    Read More

    Abcam community

    Verified customer

    Asked on Aug 29 2002

    Answer

    I'm sorry to hear that you are having trouble with ab72. Over the last couple of months, we have received two reviews from customers who are very pleased with the results of this antibody, please see the reviews section of the online datasheet for more details. Perhaps you have received a faulty vial of this antibody? We would be happy to forward a second vial or alternatively, you might like to try either of our two polyclonals to cyclin B1. (ab7957 and ab2096) Please let me know how you wish to proceed.

    Read More

    Abcam Scientific Support

    回复于 Aug 29 2002

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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