Overview

  • Product name
  • Description
    Rabbit polyclonal to CENPB
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Sheep, Hamster
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 550 to the C-terminus of Human CENPB.

    (Peptide available as ab28019.)

  • Positive control
    • This antibody gave a positive signal in the following: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate A431 (Human epithelial carcinoma cell line) Whole Cell Lysate Testis (Mouse) Tissue Lysate - normal tissue Ovary (Mouse) Tissue Lysate - normal tissue PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab25734 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
IHC-P
ICC/IF
  • Application notes
    ICC/IF: 1/200 - 1/1000. Methanol-fixed cells seem to give better results than paraformaldehyde-fixed. Please see abreview for further details.
    IHC-P: 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
    WB: Use at a concentration of 1 - 5 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function
      Interacts with centromeric heterochromatin in chromosomes and binds to a specific subset of alphoid satellite DNA, called the CENP-B box. May organize arrays of centromere satellite DNA into a higher order structure which then directs centromere formation and kinetochore assembly in mammalian chromosomes.
    • Sequence similarities
      Contains 1 HTH CENPB-type DNA-binding domain.
      Contains 1 HTH psq-type DNA-binding domain.
    • Cellular localization
      Nucleus. Chromosome > centromere.
    • Information by UniProt
    • Database links
    • Alternative names
      • CENP B antibody
      • CENP-B antibody
      • CENPB antibody
      • CENPB_HUMAN antibody
      • centromere autoantigen B antibody
      • Centromere protein B antibody
      • Centromere protein B, 80kDa antibody
      • Major centromere autoantigen B antibody
      see all

    Images

    • All lanes : Anti-CENPB antibody (ab25734) at 1 µg/ml

      Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lane 2 : Jurkat whole cell lysate (ab7899)
      Lane 3 : A431 whole cell lysate (ab7909)

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
      Developed using the ECL technique

      Predicted band size : 65 kDa
      Observed band size : 65 kDa
      Additional bands at : 74 kDa (possible cross reactivity),75 kDa (possible cross reactivity).
    • 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for CENPB (red) using ab25734 at 1/200 dilution in ICC/IF.

      The nuclear counterstain is DAPI (blue). Alpha-Tubulin is stained with an mouse monoclonal anti-alpha Tubulin antibody (green).

       

      See Abreview

    •  Image courtesy of Human Protein Atlas. ab25734 staining CENPB in Human skin, showing nuclear staining in the epidermis and dermal layer. Paraffin embedded human skin tissue was incubated with ab25734(1/100 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

      ab25734 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues. Further results for this antibody can be found at www.proteinatlas.org

    • ab25734 (1/1000) staining CENPB in HeLa cells (green). Cells were fixed with methanol and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

      See Abreview

    • ICC/IF image of ab25734 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab25734, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HEK 293 cells.

    References

    This product has been referenced in:

    See all 18 Publications for this product

    Customer reviews and Q&As

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - PBS+0.2% Triton X-100 for 30 minutes
    Specification
    HeLa
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C
    Fixative
    4% formaldehyde in PBS+0.2% Triton X-100
    Username

    Weiguo Zhang

    Verified customer

    Submitted Aug 25 2015

    Merci pour votre réponse.

    Est-ce que vous avez aussi utilisé une dilution de 1/200 pour l'anticorps primaire? Comme j'ai mentionné dans mon émail précédant, en fixation PFA, la dilution de l'anticorps...

    Read More

    Merci d'avoir pris le temps de nous expliquer un peu plus le protocole que vous avez utilisé. Je suis désolée que vous avez eu des difficultés à obtenir de bons résultats avec ab25734.

    Notre politiqu...

    Read More

    Je suis désolée d'apprendre quevotre tube d'ab25734 ne vous donne pas de bons résultats.

    Nous allons volontiers vous remplacer ce tube. Est-ce que vous pouvez nous préciser si le tube que votre collègue vous...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Lymphoblast chromosomes)
    Specification
    Lymphoblast chromosomes
    Fixative
    Carnoy (Methanol:acetic acid 3:1)
    Permeabilization
    Yes - 0,25% Triton X-100 in blocking buffer
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
    Username

    Miss. Marie-Chantal Grégoire

    Verified customer

    Submitted Sep 30 2008

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Methanol
    Permeabilization
    No
    Username

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Feb 18 2008

    Thank you for your enquiry. I have had a look at the Earnshaw publication that you highlight and I appreciate the discrepancy that you highlight. I have passed this enquiry onto my colleague who generated this datasheet and managed the antibody pro...

    Read More

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