Overview

  • Product name

    Anti-Cyclin B2/CCNB2 antibody [X29.2]
    See all Cyclin B2/CCNB2 primary antibodies
  • Description

    Mouse monoclonal [X29.2] to Cyclin B2/CCNB2
  • Host species

    Mouse
  • Specificity

    Reacts with most cyclin Bs.
  • Tested applications

    Suitable for: IHC-P, ICC/IF, WB, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Xenopus laevis
  • Immunogen

    Full length protein corresponding to Xenopus laevis Cyclin B2/CCNB2.

  • General notes

     This product was previously labelled as Cyclin B2

     

Properties

Applications

Our Abpromise guarantee covers the use of ab18250 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 Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
WB Use a concentration of 2 µg/ml. Predicted molecular weight: 45 kDa. The antibody may give a high background if the concentration of the antibody is above 2µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

Images

  • All lanes : Anti-Cyclin B2/CCNB2 antibody [X29.2] (ab18250) at 2 µg/ml

    Lane 1 : Whole cell lysate of Mitotic Hela cells (arrested by the spindle checkpoint due to depolymerization of microtubles by exposure to Nocodazole)
    Lane 2 : Whole cell lysate of Interphase Hela Cells, forced to exit mitosis by exposure to a Cdk1 inhibitor (exit from mitosis results in the degradation of Cyclin B2/CCNB2)

    Secondary
    All lanes : HRP-Conjugated Goat anti-Mouse diluted 1/20000

    Developed using the ECL technique.

    Predicted band size: 45 kDa
    Observed band size: 45 kDa
    Additional bands at: 90 kDa (possible non-specific binding)


    Exposure time: 25 minutes


    The gel was denaturing. Blocked with a 5% milk solution for 2 hours at 24°C.

    The primary antibody was diluted in 5%Milk/Tris-Buffered Saline/0.05%Tween20 and incubated for 16 hours at 4°C.

    See Abreview

  • IHC image of ab18250 staining in human cervical carcinoma 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 ab18250, 1µg/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.

  • Overlay histogram showing HeLA cells stained with ab18250 (red line). The cells were fixed with 100% 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 (ab18250, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was Mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:

  • Lin J  et al. Exportin-T promotes tumor proliferation and invasion in hepatocellular carcinoma. Mol Carcinog 58:293-304 (2019). Read more (PubMed: 30334580) »
  • Ghelli Luserna Di Rorà A  et al. Targeting WEE1 to enhance conventional therapies for acute lymphoblastic leukemia. J Hematol Oncol 11:99 (2018). Read more (PubMed: 30068368) »
See all 12 Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Application
Western blot
Sample
Xenopus laevis Cell lysate - whole cell (early embryos before MBT)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
1 cells
Specification
early embryos before MBT
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Jolanta Kisielewska

Verified customer

Submitted Jul 23 2009

Application
Western blot
Sample
Human Cell lysate - whole cell (Hela)
Gel Running Conditions
Reduced Denaturing
Loading amount
10000 cells
Treatment
Nocodazole 4 hours, Mitotic Shake-Off
Specification
Hela
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Nov 27 2007

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