Anti-CBX4 抗体 (ab4189)

Submit a review Q&A (1)References (1)
Immunocytochemistry/ Immunofluorescence - Anti-CBX4 antibody (ab4189)
  • Western blot - Anti-CBX4 antibody (ab4189)

Key features and details

  • Rabbit polyclonal to CBX4
  • Suitable for: ICC/IF, WB
  • Reacts with: Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-CBX4 antibody
    CBX4 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to CBX4

  • 由来種

    Rabbit

  • 特異性

    In immunofluoresence, this antibody detects the expected discrete nuclear structure that is termed the PcG body, corresponding to the known localisation of PC2. The antibody has been successfully used to detect FLAG-tagged transfected hPC2 (see picture). It detects a weak band that probably corresponds to endogenous PC2, however, a strong secondary band is also seen at 50kD in all cell lines thus far tested. This suggests that the antibody also reacts with another ubiquitous highly expressed protein.

  • アプリケーション

    適用あり: ICC/IF, WBmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide:

    GLQDSSTDNRAKLC

    , corresponding to amino acids 95 - 107 of Human PC2.
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • 特記事項


    PC2 and other members of the Drosophila Polycomb group gene family are part of a cellular memory system that is responsible for the inheritance of gene activity by progeny cells.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab4189 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF Use at an assay dependent dilution.
WB Use at an assay dependent dilution. Detects a band of approximately 82 kDa (predicted molecular weight: 61 kDa).

ターゲット情報

  • 機能

    E3 SUMO-protein ligase which facilitates SUMO1 conjugation by UBE2I. Component of the Polycomb group (PcG) multiprotein PRC1 complex, a complex required to maintain the transcriptionally repressive state of many genes, including Hox genes, throughout development. PcG PRC1 complex acts via chromatin remodeling and modification of histones; it mediates monoubiquitination of histone H2A 'Lys-119', rendering chromatin heritably changed in its expressibility.

  • 組織特異性

    Ubiquitous.

  • パスウェイ

    Protein modification; protein sumoylation.

  • 配列類似性

    Contains 1 chromo domain.

  • 翻訳後修飾

    Phosphorylated on Thr-497 by HIPK2 upon DNA damage; which enhances E3 SUMO-protein ligase activity and promotes sumoylation on Lys-494.

  • 細胞内局在

    Nucleus.
  • Information by UniProt

  • 参照データベース

    • 別名

      • CBX 4 antibody
      • CBX4 antibody
      • Cbx4 chromobox homolog 4 (Drosophila Pc class) antibody
      • CBX4_HUMAN antibody
      • Chromobox homolog 4 (Pc class homolog, Drosophila) antibody
      • Chromobox homolog 4 antibody
      • Chromobox protein homolog 4 antibody
      • E3 SUMO protein ligase CBX 4 antibody
      • E3 SUMO protein ligase CBX4 antibody
      • E3 SUMO-protein ligase CBX4 antibody
      • hPc 2 antibody
      • hPc2 antibody
      • NBP 16 antibody
      • NBP16 antibody
      • NS5ATP1 binding protein 16 antibody
      • Pc 2 antibody
      • Pc class 2 homolog antibody
      • Pc class homolog antibody
      • Pc class homolog Drosophila antibody
      • PC2 antibody
      • Polycomb 2 homolog antibody
      see all

    画像

    • Immunocytochemistry/ Immunofluorescence - Anti-CBX4 antibody (ab4189)
      Immunocytochemistry/ Immunofluorescence - Anti-CBX4 antibody (ab4189)This image is courtesy of Luke Hughes-Davies and Rhiannon Jade, Gurdon Institute, Cambridge, UK

      Immunofluorescent imaging of human cells (U2OS) with ab4189 reveals the expected discrete nuclear structure that is termed the PcG body corresponding to the known localisation of PC2. This corresponds to results seen in Satijn et al.

      IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.  Primary antibody used at 1/200 in 5% milk / 0.2% TWEEN for one hour, secondary antibody for 30 minutes.  All blocking and incubation steps carried out at 37 degrees C. Nuclei counterstained with Hoechst stain (blue).

    • Western blot - Anti-CBX4 antibody (ab4189)
      Western blot - Anti-CBX4 antibody (ab4189)This image is courtesy of Dr Ari Melnick, Albert Einstein College of Medicine
      All lanes : Anti-CBX4 antibody (ab4189)

      Lane 1 : HEK 293 lysate over-expressing DDDDK tagged hPC2
      Lane 2 : HEK 293 lysate

      Predicted band size: 61 kDa
      Observed band size: 75 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 30 kDa (possible non-specific secondary antibody binding), 49 kDa (possible non-specific secondary antibody binding), 65 kDa (possible non-specific secondary antibody binding)



      ab4189 specifically recognises over expressed DDDDK tagged hPC2 in HEK293 cells (lane1). The lower bands are assigned to degradation products.

    参考文献 (1)

    ab4189 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab4189 は 1 報の論文で使用されています。

    • MacPherson MJ  et al. The CTCF insulator protein is posttranslationally modified by SUMO. Mol Cell Biol 29:714-25 (2009). ICC/IF ; Human . PubMed: 19029252

    レビューと Q&A

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    Question

    BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Non-specific staining. I get mostly cytoplasmic staining rather than nuclear staining. SAMPLE U20S osteosarcoma cell line. PRIMARY ANTIBODY PC2 (ab4189) / 1:200 dilution/ 1 hr incubation. Three 10 min. washes with PBS. SECONDARY ANTIBODY Alexa Fluor Anti-Rabbit in 5% milk and 0.2% tween for 30 min. / 1:600 dilution / Three 10 min. washes with PBS ANTIBODY STORAGE CONDITIONS -20 degrees C FIXATION OF SAMPLE 4% Paraformaldehyde. PERMEABILIZATION STEP 0.5% Triton for 5 min. BLOCKING CONDITIONS 1 hr. with 5% milk/ 0.2% tween. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

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    Answer

    I'm sorry to hear you are having problems with ab4189, I would like to suggest a few modifications to your protocol: -try a different fixation step, I would suggest a 10min acetone or methanol (ice cold) fixation, it can be a very good fixative in ICC and can also help the permeabilisation to promote penetration of the antibody into the nucleus. -block the cells in 5%BSA in PBST (triton 0.3%) 30min-1hr -incubate the primary antibody overnight at a 1:200 dilution in PBST (triton 0.3%) -make sure the cytoplasmic staining you see is not caused by the secondary by omitting the primary antibody. The Alexa fluor antibodies are very powerful, you may wish to try more dilute antibody (1:1000, 1:2000) and incubate longer (1hr). I hope these suggestions will help, please let me know how you get on and do not hesitate to contact us again if you still have problems,

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