Anti-Cyclin B2抗体[X29.2] (ab18250)

概述

  • 产品名称Anti-Cyclin B2抗体[X29.2]
    参阅全部 Cyclin B2 一抗
  • 描述
    小鼠单克隆抗体[X29.2] to Cyclin B2
  • 特异性Reacts with most cyclin Bs.
  • 经测试应用适用于: IHC-P, ICC/IF, WB, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Xenopus laevis
  • 免疫原

    Full length protein (Xenopus laevis).

性能

应用

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.

应用 Ab评论 说明
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.

靶标

Anti-Cyclin B2 antibody [X29.2] 图像

  • All lanes : Anti-Cyclin B2 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)

    Secondary
    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

    This image is courtesy of an anonymous Abreview

    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.

Anti-Cyclin B2 antibody [X29.2] (ab18250)参考文献

This product has been referenced in:
  • Davies C  et al. Murine norovirus replication induces G0/G1 cell cycle arrest in asynchronously growing cells. J Virol 89:6057-66 (2015). Read more (PubMed: 25810556) »
  • Dupré A  et al. Phosphorylation of ARPP19 by protein kinase A prevents meiosis resumption in Xenopus oocytes. Nat Commun 5:3318 (2014). Read more (PubMed: 24525567) »

See all 5 Publications for this product

Product Wall

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 yo...

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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 i...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Xenopus laevis Cell lysate - whole cell (early embryos before MBT)
Loading amount 1 cells
Specification early embryos before MBT
Gel Running Conditions Reduced Denaturing (4-12%)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Jolanta Kisielewska

Verified customer

提交于 Jul 23 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Hela)
Loading amount 10000 cells
Specification Hela
Treatment Nocodazole 4 hours, Mitotic Shake-Off
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

提交于 Nov 27 2007

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 t...

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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 ...

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