Overview

  • Product name

    Anti-Cyclin B1 antibody [V152] (HRP)
    See all Cyclin B1 primary antibodies
  • Description

    Mouse monoclonal [V152] to Cyclin B1 (HRP)
  • Host species

    Mouse
  • Conjugation

    HRP
  • Tested applications

    Suitable for: IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Fusion protein corresponding to Hamster Cyclin B1.

  • Positive control

    • In Western Blot, this antibody gave a positive signal in the following whole cell lysates: K562; Jurkat; HEK293. IHC-P: FFPE human tonsil (normal) tissue sections.
  • General notes

     

     

Properties

Applications

Our Abpromise guarantee covers the use of ab201853 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
IHC-P 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/5000. Detects a band of approximately 55 kDa (predicted molecular weight: 48 kDa).

Abcam recommends using 5% Milk as the blocking agent.

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.
  • Information by UniProt
  • Database links

  • 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

  • All lanes : Anti-Cyclin B1 antibody [V152] (HRP) (ab201853) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : CCNB1 (Cyclin B1) knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 48 kDa
    Observed band size: 51 kDa
    why is the actual band size different from the predicted?


    Exposure time: 12 minutes


    ab201853 was shown to specifically react with Cyclin B1 in wild-type HAP1 cells as signal was lost in CCNB1 (Cyclin B1) knockout cells. Wild-type and CCNB1 (Cyclin B1) knockout samples were subjected to SDS-PAGE. Ab201853 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

  • IHC image of Cyclin B1 staining in a section of formalin-fixed paraffin-embedded normal human tonsil tissue*, 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 20mins. The section was then incubated with ab201853, 1/250 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative 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

  • All lanes : Anti-Cyclin B1 antibody [V152] (HRP) (ab201853) at 1/5000 dilution

    Lane 1 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 48 kDa
    Observed band size: 51 kDa why is the actual band size different from the predicted?
    Additional bands at: 36 kDa (possible non-specific binding)


    Exposure time: 20 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab201853 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

References

ab201853 has not yet been referenced specifically in any publications.

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