Invitrogen

Ki-67 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated. View Details

4 Published Figures

9 References

View all (119) Ki-67 antibodies

Datasheet

Questions & Answers

Datasheet

Questions & Answers

Product Image Gallery

Ki-67 Antibody in Immunofluorescence (IF) — Immunofluorescence analysis of Ki67 was performed using 70% confluent log phase HeLa cells serum starved for 36 hours followed by serum release for 6 hrs. The cells were fixed with 4% Paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 10 minutes at room temperature. The cells were labeled with Ki67 Polyclonal Antibody (Product # PA1-21520) at 10ug/ml in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034), (1:2000 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415 , 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents the serum starved cells with reduced signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Ki-67 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of Ki67 in formalin-fixed, paraffin-embedded human tonsil tissue using a Ki67 polyclonal antibody (Product # PA1-21520) and detected using ABC method with AEC chromogen.

Ki-67 Antibody in Immunohistochemistry (IHC) — Immunohistochemical analysis of Ki67 in HT-29 xenograft tumor in mouse using a Ki67 polyclonal antibody (Product # PA1-21520) at a dilution of 5 µg/mL for 30 min at room temperature, following a 20x pretreatment with a citrate buffer, pH 6.0.

Ki-67 Antibody in Cell treatment — Detection of altered expression of the target protein by cell treatment demonstrates antibody specificity. Immunofluorescence analysis of Ki67 using Ki67 Polyclonal Antibody (Product # PA1-21520), shows decreased expression of Ki67 in HeLa cell line upon serum starvation for 36 hours. Serum starvation followed by serum release for 6 hours resulted in increase in Ki67 expression in the nucleus of HeLa cells. {TM}

Ki-67 Antibody — Fig. 1. GA treatment compromises the viability of breast cancer cells. (A) Expression of SUMO-conjugated proteins (nSUMO1) in different molecular subtypes of breast cancer cells: luminal (MCF7), triple-negative (MDA-MB-231) and HER2+ (BT474) treated with GA (10 uM) for 24 h and analysed by immunoblotting (12% acrylamide gel). (B) MCF7, MDA-MB-231 and BT474 breast cancer cell lines were treated with various concentrations of GA and their cell viability was assessed in an MTT assay: * P <=0.001; ** P <=0.0001; n.s., not significant versus non-treated cells (two-way ANOVA). Turkey's multiple comparisons shows statistical significant differences in all conditions apart from 10 uM GA at 24 h. (C) Number of MDA-MB-231 cells after 2 days of treatment with different concentrations of GA relative to the control ( t =0 h treatment). (D) Representative image of immunohistochemical evaluation of Ki-67 expression in MDA-MB-231 tumour cells and (E) the percentage of Ki-67-positive cells. (F) Expression of SUMO-conjugated proteins (nSUMO1) in MDA-MB-231 cells exposed to GA (10 uM) or 2-D08 (20 uM) for different times and analysed by immunoblotting (12% acrylamide gel). (G) Number of MDA-MB-231 cells after 2 days of treatment with different concentrations of 2-D08 relative to the controls ( t =0 h treatment). (H) Percentage of Ki-67-positive MDA-MB-231 cells after 2-D08 treatment for 24 h. (I) SUMO-conjugated proteins in MDA-MB-231 control and SUMO1-depleted cells analysed by immunoblotting

Product Details

Applications

Tested Dilution

Publications

Immunocytochemistry (ICC)

10 µg/mL

View 2 publications 2 publications

Immunofluorescence (IF)

10 µg/mL

View 1 publication 1 publication

Immunohistochemistry (IHC)

Assay Dependent

View 4 publications 4 publications

Immunohistochemistry (Paraffin) (IHC (P))

10-20 µg/mL

View 1 publication 1 publication

Western Blot (WB)

View 1 publication 1 publication

Product Specifications

Species

Human, Mouse, Rat, Zebrafish

Published species

Human, Mouse

Expression System

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide from the human Ki-67 antigen.

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

10mM PBS, pH 7.4, with 0.2% BSA

Contains

0.1% sodium azide

Storage conditions

4° C, do not freeze

RRID

AB_559806

Product Specific Information

Recommended positive controls: Tonsil or Breast carcinoma.

Staining of formalin/paraffin tissues requires boiling tissue sections in 10 mM citrate buffer, pH6.0, for 10-20 minutes, followed by coolong at room temperature for 20 minutes.

Target Information

Ki-67 is a nuclear protein that is expressed during various stages in the cell cycle, particularly during late G1, S, G2, and M phases. The protein has a forkhead associated domain (FHA) through which it associates with euchromatin at the perichromosomal layer, the centromeric heterochromatin, and the nucleolus. Ki-67 is shown to have a cell cycle dependent topographical distribution with perinucleolar expression at G1, expression in the nuclear matrix at G2, and expression on the chromosomes during M phase. Ki-67 is commonly used as a proliferation marker because it is not detected in G0 cells, but increases steadily from G1 through mitosis. Ki-67 antibodies are useful in establishing the cell growing fraction in neoplasms. In neoplastic tissues, the prognostic value is comparable to the tritiated thymidine-labelling index. The correlation between low Ki-67 index and histologically low-grade tumors is strong. Ki-67 is routinely used as a neuronal marker of cell cycling and proliferation.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: Antigen identified by monoclonal antibody Ki-67; Antigen identified by monoclonal antibody Ki-67 homolog; Antigen KI-67; Antigen KI-67 homolog; antigen KI-67-like; Proliferation marker protein Ki-67; proliferation-related Ki-67 antigen; protein phosphatase 1, regulatory subunit 105; RP11-380J17.2

Gene Aliases: cb31; D630048A14Rik; Ki-67; Ki67; KIA; MIB-; MIB-1; MKI67; PPP1R105; sb:cb31; si:ch211-250b22.7; wu:fa11g09; wu:fb57a07; wu:fi14e05

UniProt ID: (Human) P46013, (Mouse) E9PVX6

Entrez Gene ID: (Human) 4288, (Rat) 291234, (Mouse) 17345, (Zebrafish) 564521

Function(s)

DNA metabolic process meiotic nuclear division cell proliferation response to organic cyclic compound hyaluronan metabolic process organ regeneration cellular response to heat hepatocyte proliferation cholangiocyte proliferation

Process(es)

protein binding ATP binding protein C-terminus binding poly(A) RNA binding molecular_function