Validated using a knockout cell line
Recombinant
RabMAb

Anti-Cyclin B1 antibody [Y106] - BSA and Azide free (ab156447)

Overview

  • Product name
    Anti-Cyclin B1 antibody [Y106] - BSA and Azide free
    See all Cyclin B1 primary antibodies
  • Description
    Rabbit monoclonal [Y106] to Cyclin B1 - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Cyclin B1 aa 400-500 (C terminal). The exact sequence is proprietary.

  • General notes

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    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 a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab156447 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
WB Use at an assay dependent concentration. Predicted molecular weight: 58 kDa.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.

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

  • 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 secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32053).

  • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Cyclin B1 with purified ab32053 at 1:400 dilution (1 µg/ml) (red). 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). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32053).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32053).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32053).

References

ab156447 has not yet been referenced specifically in any publications.

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