Anti-Cyclin B2/CCNB2 antibody [X29.2] (ab18250)

Reviews (3)Q&A (4)References (16)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B2/CCNB2 antibody [X29.2] (ab18250)
  • Flow Cytometry - Anti-Cyclin B2/CCNB2 antibody [X29.2] (ab18250)

Key features and details

  • Mouse monoclonal [X29.2] to Cyclin B2/CCNB2
  • Suitable for: IHC-P, Flow Cyt
  • Reacts with: Human
  • Isotype: IgG1

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, Flow Cytmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • General notes

     This product was previously labelled as Cyclin B2

     

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

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.
Flow Cyt
Use 1µg for 106 cells.

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

 
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.
Flow Cyt
Use 1µg for 106 cells.

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

 

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.
  • Information by UniProt

  • Database links

    • Alternative names

      • ccnb2 antibody
      • CCNB2_HUMAN antibody
      • CycB2 antibody
      • Cyclin B2 antibody
      • G2 mitotic specific cyclin B2 antibody
      • G2/mitotic specific cyclin B2 antibody
      • G2/mitotic-specific cyclin-B2 antibody
      • HsT17299 antibody
      • MGC108931 antibody
      • MGC140694 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B2/CCNB2 antibody [X29.2] (ab18250)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B2/CCNB2 antibody [X29.2] (ab18250)
      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.

    • Flow Cytometry - Anti-Cyclin B2/CCNB2 antibody [X29.2] (ab18250)
      Flow Cytometry - Anti-Cyclin B2/CCNB2 antibody [X29.2] (ab18250)
      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 (16)

    Publishing research using ab18250? Please let us know so that we can cite the reference in this datasheet.

    ab18250 has been referenced in 16 publications.

    • Huang S  et al. Obg-like ATPase 1 (OLA1) overexpression predicts poor prognosis and promotes tumor progression by regulating P21/CDK2 in hepatocellular carcinoma. Aging (Albany NY) 12:3025-3041 (2020). PubMed: 32045367
    • Wang X  et al. Effect of miR-205 on proliferation and migration of thyroid cancer cells by targeting CCNB2 and the mechanism. Oncol Lett 19:2568-2574 (2020). PubMed: 32194761
    • Lin J  et al. Exportin-T promotes tumor proliferation and invasion in hepatocellular carcinoma. Mol Carcinog 58:293-304 (2019). PubMed: 30334580
    • Yi ZY  et al. PKCß1 regulates meiotic cell cycle in mouse oocyte. Cell Cycle 18:395-412 (2019). PubMed: 30730241
    • Ghelli Luserna Di Rorà A  et al. Synergism Through WEE1 and CHK1 Inhibition in Acute Lymphoblastic Leukemia. Cancers (Basel) 11:N/A (2019). PubMed: 31717700
    View all Publications for this product

    Customer reviews and Q&As

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    1-7 of 7 Abreviews or Q&A

    Western blot abreview for Anti-Cyclin B2/CCNB2 antibody [X29.2]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Cancer cells - MDA/MB731)
    Gel Running Conditions
    Reduced Denaturing (4-16%)
    Loading amount
    30 µg
    Treatment
    Local anesthetics 50uM
    Specification
    Cancer cells - MDA/MB731
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

    Dr. Michele D'angelo

    Verified customer

    Submitted Jan 29 2020

    Question

    Thanks for review this data. We have rerun the test again according to your below recommendations, we were still unable to achieve a good outcome. I am attaching the data with email for your review again. we hope to hear of your advise, and whether a different lot might be accessible.

    We apologize for any inconveniences.

    Read More
    Answer

    Thank you for your reply. I am sorry that the antibodies are still not producing good results in WB. We do not currently have new lots available for either product, so I have issued you a refund (Credit Note: ***). To redeem this refund, please have your purchasing agent contact our accounting department at us.credits@abcam.com. Please let me know if you have any further questions.

    Read More

    Question

    Attached please find the antibody ab18250 & ab75743wb questionnaire answer sheets. We did not observe an optimal results over these two antibodies. we would look forward to any suggestion or advice you may have.

    Read More
    Answer

    Thank you for contacting us. I am sorry that these antibodies are giving weak signal in your samples.

    For Cyclin B2, I would expect ab18250 to give good staining using the protocol you sent, however this protein is not very highly expressed in normal or cancerous liver. You may find better results testing a positive control such as bone marrow or HeLa cells.

    The antibody ab75742 is phospho-specific, so I would not recommend using milk as a blocking agent. Milk contains endogenous phosphatases which can cleave the phosphorylations of interest and results in significantly reduced signal. Switching to BSA may help to improve the strength of your band. Have you done any treatment as a positive control to ensure that this particular phosphorylation is expressed?

    I hope this helps, if not, please let me know and I will be happy to help you further.

    Read More

    Western blot abreview for Anti-Cyclin B2/CCNB2 antibody [X29.2]

     Average
    Abreviews
    abreview image
    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
    Read More

    Dr. Jolanta Kisielewska

    Verified customer

    Submitted Jul 23 2009

    Western blot abreview for Anti-Cyclin B2/CCNB2 antibody [X29.2]

     Good
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 27 2007

    Question

    I' m sending the informations that can be useful to receive suggestions about 18250 Ab. Blocking solution prepared: 3% BSA in PBS 1X Antibody dilution utilized: 1/1000 in blocking solution Waiting your reply, all my best regards.

    Read More
    Answer

    Thank you for your e-mail. I would recommend changing: - the blocking buffer to TBST (tris HCl (pH7.6) with Tween20 0.1%) and trying 5%BSA (1hr) or 5% milk (1hr) -the antibody dilution buffer to TBST only and try also 0.2% milk in the TBST. I think one of those options will work for you. Here is my recipe for the TBST buffer: • TBS 10x (concentrated TBS) -24.23g Trizma HCl -80.06g NaCl Mix in 800ml ultra pure water. pH to 7.6 with pure HCl. Top up to 1L. • TBST For 1L: 100ml of TBS 10x + 900ml ultra pure water + 1ml Tween 20 Please let me know if you require further assistance,

    Read More

    Question

    Abcam Order Number: 73632 Ab18250 Batch number: 144180 This is to inform you on the reactivity of ab18250 raised against Xenopus Cyclin B2. Initial attempts to recognize cyclin B2 from total cell extracts deriving from Rana temporaria tissues by immunoblot failed. Therefore, we tested the antibody against a positive control represented by metaphase II blocked Xenopus eggs. According to Gautier and Maller (EMBO J. 1991 Jan;10(1):177-82) these extracts should contain plently of cyclin B2. However, as you can see from the attached image, we were unable to detect any cross-reacting band on WB at the expected molecular wiight, i.e. about 45 kDa according to the Ab18250 data sheet. On the contrary, we got multiple and not-specific bands. Could you please help us to better settle down our immunoblot? Do you advice a different positive control? If no solutions are available should we replace this antibody with a different one's? We are waiting for an answer at your convenience. Sincerely yours,

    Read More
    Answer

    The originating scientist has got back to me about your query. He says that he has found that the antibody may give a high background in an immunoblot if the concentration of the antibody is above 2µg/ml. He says that you should optimise using a range of dilutions and may need to change blocking buffer. I am still waiting for the positive control information,my apologies for the delay,

    Read More

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