Overview

  • Product nameAnti-Cyclin B1 antibody [V152]See all Cyclin B1 primary antibodies ...
  • Description
    Mouse monoclonal [V152] to Cyclin B1
  • SpecificityCyclin B1 expression is restricted to a specific short period of the cell cycle with cyclin B1 expression detected earlier and peaking in concentration before cyclin B2 expression.
  • Tested applicationsSDS-PAGE, ICC/IF, IHC (PFA fixed), Flow Cyt, WB, IHC-FoFr, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    His-tagged Hamster Cyclin B1 expressed in bacteria, harvested from inclusion bodies, extracted with 6M guanidine HCl and purified on Nickel beads.

  • Positive control
    • In Western Blot, this antibody gave a positive signal in the following whole cell lysates: HeLa; Daudi; K562; Jurkat; HEK293. In IHC, positive staining was observed in a Human Normal Tonsil formalin fixed paraffin embedded tissue section.

Properties

Applications

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.

Application Abreviews Notes
SDS-PAGE 1/1500.
ICC/IF Use at an assay dependent concentration.
IHC (PFA fixed) 1/400.
Flow Cyt Use at an assay dependent concentration.
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-FoFr Use a concentration of 1 - 2 µg/ml.
IHC-P Use a concentration of 5 µg/ml.
IHC-Fr Use a concentration of 1 - 2 µg/ml.
  • Application notesIs unsuitable for IP.
  • Target

    • FunctionEssential for the control of the cell cycle at the G2/M (mitosis) transition.
    • Sequence similaritiesBelongs to the cyclin family. Cyclin AB subfamily.
    • Developmental stageAccumulates 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 localizationCytoplasm. Nucleus. Cytoplasm > cytoskeleton > centrosome.
    • Information by UniProt
    • Database links
    • Alternative names
      • CCNB 1 antibody
      • CCNB antibody
      • ccnb1 antibody
      • CCNB1_HUMAN antibody
      • G2 mitotic specific cyclin B1 antibody
      • G2/mitotic-specific cyclin-B1 antibody
      see all

    Anti-Cyclin B1 antibody [V152] images

    • 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 (Human Burkitt's lymphoma cell line) 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 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      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
    • IHC image of ab72 staining in Human Normal Tonsil formalin fixed paraffin embedded tissue 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 (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab72, 5µg/ml, 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.

      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.



    • Predicted band size : 48 kDa


      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 whole cell lysate loaded at 25ug per lane. Ab72 used at 1/500 in reducing conditions in conjunction with goat anti mouse (HRP) 1/20000.

      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

    References for Anti-Cyclin B1 antibody [V152] (ab72)

    This product has been referenced in:
    • Nabti I  et al. Dual-mode regulation of the APC/C by CDK1 and MAPK controls meiosis I progression and fidelity. J Cell Biol 204:891-900 (2014). WB . Read more (PubMed: 24637322) »
    • Luo Y  et al. Human parvovirus B19 infection causes cell cycle arrest of human erythroid progenitors at late S phase that favors viral DNA replication. J Virol 87:12766-75 (2013). Human . Read more (PubMed: 24049177) »

    See all 32 Publications for this product

    Product Wall

    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
    Username

    Abcam user community

    Verified customer

    Submitted Nov 01 2013

    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
    Username

    Dr. Brandon White

    Verified customer

    Submitted Aug 29 2012

    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
    Username

    Abcam user community

    Verified customer

    Submitted Aug 17 2011

    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
    Username

    Abcam user community

    Verified customer

    Submitted Nov 18 2008

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (HeLa)
    Specification HeLa
    Fixative Methanol
    Permeabilization No
    Username

    Dr. Kirk McManus

    Verified customer

    Submitted May 12 2008

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

    Abcam user community

    Verified customer

    Submitted Sep 12 2005

    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...

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"