Invitrogen

Ki-67 Monoclonal Antibody (SolA15), eBioscience™

Advanced Verification

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

40 Published Figures

154 References

View all (119) Ki-67 antibodies

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Datasheet

Questions & Answers

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Ki-67 Antibody in Immunofluorescence (IF) — Immunofluorescence analysis of Ki67 was performed using 70% confluent log phase HeLa cells serum starved for 36 Hrs 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 Ki-67 Monoclonal Antibody (SolA15), eBioscience™ (Product # 14-5698-82) at 5 µg/ml in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A1-1006, 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 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 Immunocytochemistry (ICC) — Immunocytochemistry on fixed and permeabilized C2C12 cells using 5 µg/mL of Rat IgG2a Isotype Control (left) or 5 µg/mL of Anti-Mouse/Rat Ki-67 Purified (right) followed by Anti-Rat TRITC. Nuclei are stained with DAPI. Co-expression of Ki-67 and DAPI appears pink.

Ki-67 Antibody in Immunohistochemistry (IHC) — Immunofluorescent analysis was performed with formalin-fixed paraffin embedded mouse stomach tumor sections. The slides were de-paraffinization and undergo antigen retrieval in TE pH9. The sections were blocked with 5% normal donkey serum in PBS for an hour, then incubated with the Rat anti-Ki67 antibody (Product # 14-5698-80) diluted 1:200 overnight. The slides were then washed three times with TBS, and incubated with the CY3 conjugated donkey anti-rat Cy3 secondary antibody for 1.5 hours. Finally the slides were counterstained with Hoechst (Product # 62249) diluted 1:5000 before mounting in Prolong Gold anti-fad mounting media (Product # P36931). Images were acquired using a confocal microscope. The positive and negative images show sections with and without the primary antibody respectively, acquired with the same imaging parameters. Data courtesy of Antibody Data Exchange Program.

Ki-67 Antibody in Cell treatment — Immunofluorescence analysis of Ki67 was performed using 70% confluent log phase HeLa cells serum starved for 36 Hrs 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 Ki-67 Monoclonal Antibody (SolA15), eBioscience™ (Product # 14-5698-82) at 5 µg/ml in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (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 Cytoplasmic localization. Panel e represents HeLa cells serum starved for 36 Hrs with reduced signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification. {TM}

Ki-67 Antibody — Figure 3 Follicle-like structures of SPMS brains are devoid of FOXP3 expression, but exhibit NFATc1 + cells. (A) Representative meningeal follicle-like structure of SPMS spinal cord, which was screened based on H and E staining and characterized by >60 lymphocytes (score 3), detection of Ki67 + , CD35 + /CD21 + , and CD138 + cells on serial sections, termed F+. F-, if no or not all criteria were fulfilled. (B) Follicle-like structures could be characterized by CXCR5 + , (C) but not by FOXP3 + cells. (D) NFATc1 + cells were present in follicle-like structures. (E) IHC stainings of Ki67, CD35, CD21, BCL-6, CXCR5, and NFATc1 were quantified as frequency of total cells in follicle-like structures (F+) and less defined infiltrates (F-). Ki67 : F-, M = 1.52, SD = 3.73, n = 45; F+, M = 1.89, SD = 3.55, n = 38; Mann Whitney test, U = 734.0, p = 0.161. CD35 : F-, M = 15.41, SD = 22.95, n = 22; F+, M = 19.05, SD = 14.75, n = 21; Mann Whitney test, U = 159.0, p = 0.081. CD21 : F-, M = 5.68, SD = 10.63, n = 21; F+, M = 3.51, SD = 5.60, n = 20; Mann Whitney test, U = 183.5, p = 0.488. BCL-6 : F-, M = 0.32, SD = 1.08, n = 44; F+, M = 0.49, SD = 1.62, n = 38; Mann Whitney test, U = 803.0, p = 0.642. CXCR5 : F-, M = 14.72, SD = 17.24, n = 30; F+, M = 20.11, SD = 15.73, n = 29; Mann Whitney test, U = 319.0, p = 0.080. NFATc1 : F-, M = 7.15, SD = 11.68, n = 28; F+, M = 16.36, SD = 21.46, n = 31; Mann Whitney test, U = 307.0, p = 0.043. Scale bars A-D indicate 100 mum. *, p < 0.05.

Product Details

Applications

Tested Dilution

Publications

Immunocytochemistry (ICC)

5 µg/mL

View 6 publications 6 publications

Immunofluorescence (IF)

5 µg/mL

View 46 publications 46 publications

Immunohistochemistry (Frozen) (IHC (F))

5 µg/mL

View 4 publications 4 publications

Immunohistochemistry (Paraffin) (IHC (P))

2.5-5 µg/mL

View 8 publications 8 publications

Flow Cytometry (Flow)

View 53 publications 53 publications

Immunohistochemistry (IHC)

View 36 publications 36 publications

Western Blot (WB)

View 1 publication 1 publication

Product Specifications

Species

Dog, Cynomolgus Monkey, Human, Mouse, Non-human primate, Rat

Published species

Artificial Control, Fish, Human, Mouse, Rat

Expression System

Rat / IgG2a, kappa

Class

Monoclonal

Type

Antibody

Clone

SolA15

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4° C

RRID

AB_10854564

Product Specific Information

Description: The monoclonal antibody SolA15 recognizes mouse and rat Ki-67, a 300 kDa nuclear protein. Ki-67 is present during all active phases of the cell cycle (G1, S, G2, and mitosis), but is absent from resting cells (G0). Ki-67 is detected within the nucleus during interphase but redistributes to the chromosomes during mitosis. Ki-67 is used as a marker for determining the growth fraction of a given population of cells. In studies of tumor cells, the "Ki-67 labeling index" refers to the number of Ki-67 positive cells within the population and this is used to predict outcome of particular cancer types. Ki-67 has been shown to interact with the DNA-bound protein chromobox protein homolog 3 (CBX3) (heterochromatin).

The SolA15 antibody also recognizes human, non-human primate and canine Ki-67.

Applications Reported: This SolA15 antibody has been reported for use in immunohistochemical staining of formalin-fixed paraffin embedded (FFPE) tissue sections and frozen (paraformaldehyde fixed) sections and immunocytochemistry (ICC).

Applications Tested: This SolA15 antibody has been tested by immunohistochemistry on formalin-fixed paraffin embedded mouse skin with high or low pH antigen retrieval and immunocytochemistry on fixed and permeabilized C2C12 cells at less than or equal to 5 µg/mL. It is recommended that this antibody be carefully titrated for optimal performance in the assay of interest.

Purity: Greater than 90%, as determined by SDS-PAGE.

Aggregation: Less than 10%, as determined by HPLC.

Filtration: 0.2 µm post-manufacturing filtered.

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; Proliferation marker protein Ki-67; proliferation-related Ki-67 antigen; protein phosphatase 1, regulatory subunit 105; RP11-380J17.2; unnamed protein product

Gene Aliases: D630048A14Rik; Ki-67; Ki67; KIA; MIB-; MIB-1; MKI67; PPP1R105

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

Entrez Gene ID: (Dog) 100686578, (Human) 4288, (Cynomolgus Monkey) 102135895, (Rat) 291234, (Mouse) 17345

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