製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to Ki67
  • アプリケーション
    適用あり: ICC/IF, ICC, IHC-P, IHC-Frmore details
  • 種交差性
    交差種: Human, African green monkey
  • 免疫原

    Synthetic peptide corresponding to Human Ki67. Immunogen is from the 62 base pair region of the human Ki-67 antigen.
    Database link: P46013

  • ポジティブ・コントロール
    • Tonsil.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab833 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 1/400. PubMed: 18635871
ICC Use at an assay dependent concentration.
IHC-P 1/25 - 1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. We recommend using an ABC system for detection of this antibody.
IHC-Fr 1/25 - 1/50. 1/25 - 1/50. We suggest an incubation period of 30 minutes at room temperature and to use DAB to stain the protein.

ターゲット情報

  • 機能
    Required to maintain individual mitotic chromosomes dispersed in the cytoplasm following nuclear envelope disassembly (PubMed:27362226). Associates with the surface of the mitotic chromosome, the perichromosomal layer, and covers a substantial fraction of the chromosome surface (PubMed:27362226). Prevents chromosomes from collapsing into a single chromatin mass by forming a steric and electrostatic charge barrier: the protein has a high net electrical charge and acts as a surfactant, dispersing chromosomes and enabling independent chromosome motility (PubMed:27362226). Binds DNA, with a preference for supercoiled DNA and AT-rich DNA (PubMed:10878551). Does not contribute to the internal structure of mitotic chromosomes (By similarity). May play a role in chromatin organization (PubMed:24867636). It is however unclear whether it plays a direct role in chromatin organization or whether it is an indirect consequence of its function in maintaining mitotic chromosomes dispersed.
  • 配列類似性
    Contains 1 FHA domain.
    Contains 16 K167R repeats.
    Contains 1 PP1-binding domain.
  • 発生段階
    Expression occurs preferentially during late G1, S, G2 and M phases of the cell cycle, while in cells in G0 phase the antigen cannot be detected (at protein level) (PubMed:6206131). Present at highest level in G2 phase and during mitosis (at protein level). In interphase, forms fiber-like structures in fibrillarin-deficient regions surrounding nucleoli (PubMed:2674163, PubMed:8799815).
  • 翻訳後修飾
    Phosphorylated. Hyperphosphorylated in mitosis (PubMed:10502411, PubMed:10653604). Hyperphosphorylated form does not bind DNA.
  • 細胞内局在
    Chromosome. Nucleus. Nucleus, nucleolus. Associates with the surface of the mitotic chromosome, the perichromosomal layer, and covers a substantial fraction of the mitotic chromosome surface (PubMed:27362226). Associates with satellite DNA in G1 phase (PubMed:9510506). Binds tightly to chromatin in interphase, chromatin-binding decreases in mitosis when it associates with the surface of the condensed chromosomes (PubMed:15896774, PubMed:22002106). Predominantly localized in the G1 phase in the perinucleolar region, in the later phases it is also detected throughout the nuclear interior, being predominantly localized in the nuclear matrix (PubMed:22002106).
  • Information by UniProt
  • 参照データベース
  • 別名
    • Antigen identified by monoclonal antibody Ki 67 antibody
    • Antigen identified by monoclonal antibody Ki-67 antibody
    • Antigen KI-67 antibody
    • Antigen KI67 antibody
    • Antigen Ki67 antibody
    • KI67_HUMAN antibody
    • KIA antibody
    • Marker of proliferation Ki-67 antibody
    • MIB 1 antibody
    • MIB antibody
    • MKI67 antibody
    • PPP1R105 antibody
    • Proliferation marker protein Ki-67 antibody
    • Proliferation related Ki 67 antigen antibody
    • Protein phosphatase 1 regulatory subunit 105 antibody
    • RP11-380J17.2 antibody
    see all

画像

  • HT-29 xenograft tumor in mouse 20x pretreatment w/ citrate pH 6.0 using anti-Ki67 antibody ab833 at 5 ug/mL for 30 min at RT.
  • Immunohistochemical analysis of Human skin tissue, staining Ki67 (red) with ab833.

    Antigen retrieval was performed by heat mediation in a EDTA/Tris buffer (pH 8.0). Sections were blocked with 10% NGS for 1 hour at room temperature, followed by incubation with primary antibody (1/25). An AlexFluor®546 anti-rabbit IgG (1/500) was used as the secondary antibody.
  • Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling Ki67 with ab833 at 1/50 dilution.

    HeLa cells are the classic example of an immortalized human epithelial cell line from a fatal cervical carcinoma transformed by human papillomavirus 18 (HPV18) that grown as adherent cells layer and maintain contact inhibition in vitro. 2% paraformaldehyde for 15 min was used to fix the cells, and permealized the cells with Triton-X100 (0.1%) for 10 min and then incubated cells with primary antibody Ki-67 (1/50) for overnight at 4 degree centigrade. Image was taken using a confocal laser scanning microscope that equipped with an additional differential Interference Contrast (DIC) mode. Image here demonstrate DAPI (blue-nuclear stain, upper left panel), Ki-67 (Green, upper right panel, shows nuclear localization in actively dividing cells), Hela cells in culture-DIC (phase) image (lower left panel) and superimpose image (lower right panel).   The expression of the human Ki-67 protein is strictly associated with cell proliferation. During interphase, the antigen is exclusively detected as several prominent specks within the nucleus and a weak finely dispersed staining throughout the nucleoplasm, whereas in mitosis most of the protein is relocated to the surface of the chromosomes. The fact that the Ki-67 protein is present during all active phases of the cell cycle [G(1), S, G(2), and mitosis], but is absent from resting cells [G(0)], makes it an excellent marker for determining the so-called Ki-67 staining index (Ki67-SI) of a given cell population.

    See Abreview

  • ab833 - immunohistochemistry

    Formalin fixed paraffin embedded human tonsil stained with Ki-67, using ABC and AEC chromogen.

参考文献

This product has been referenced in:
  • Siegfried JM  et al. Interaction between the estrogen receptor and fibroblast growth factor receptor pathways in non-small cell lung cancer. Oncotarget 8:24063-24076 (2017). IHC ; Mouse, Human . Read more (PubMed: 28445992) »
  • Watanabe A  et al. Stathmin 1 promotes the proliferation and malignant transformation of pancreatic intraductal papillary mucinous neoplasms. Oncol Lett 13:1783-1788 (2017). IHC-P ; Human . Read more (PubMed: 28454324) »

See all 100 Publications for this product

レビューと Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (liver)
Permeabilization
Yes - 0.2% Triton x100
Specification
liver
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 33% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Oct 17 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Marmoset (common) Tissue sections (Colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Colon
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative
Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

投稿 Sep 25 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Tonsil
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative
Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

投稿 Aug 20 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (prostate cancer)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 0.05% trypsin for 10 min at 37°C
Permeabilization
No
Specification
prostate cancer
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

投稿 Jul 21 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Sample
African Green Monkey Cell (Cos7 Monkey Kidney Fibroblast)
Specification
Cos7 Monkey Kidney Fibroblast
Permeabilization
Yes - .3% Triton X100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Jun 13 2013

>As I mentioned previously, I would recommend running a no primary antibody control to ensure that the diffuse nuclear staining cannot be attributed to non-specific binding. If the no primary antibody control is clear, then I would conclude thatmost of...

Read More

Ki67 will be localized in the perinucleolar region in the G1 phase and in the nuclear matrix (your hazy nuclear stain) in the later S, G2, and M phases. During G0, Ki67 will not be detected. If you are looking to quantify interphase / division vs quies...

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Adult skin)
Specification
Adult skin
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric buffer, pH 6
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Abcam user community

Verified customer

投稿 Apr 06 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver)
Specification
Liver
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 May 20 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Human Cultured Cells (retinoblastoma cells)
Specification
retinoblastoma cells
Fixative
Cytospin solution: methanol, acetone, glycerine
Permeabilization
Yes - PBS-Tween
Username

Abcam user community

Verified customer

投稿 Apr 14 2009

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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