Invitrogen

Ki-67 Monoclonal Antibody (20Raj1), eBioscience™

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

16 Published Figures

21 References

View all (175) Ki-67 antibodies

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Ki-67 Antibody in Immunocytochemistry (ICC/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 (20Raj1), eBioscience™ (Product # 14-5699-82) at 1 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 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 Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry on formalin-fixed paraffin embedded human skin using 1 µg/mL of Anti-Human Ki-67 Purified (bottom) or Mouse IgG1 kappa Isotype Control Purified (Product # 14-4714-82) (inset, bottom) followed by Anti-Mouse IgG Biotin, and DAB visualization.Nuclei are counterstained with hematoxylin.

Ki-67 Antibody in Immunohistochemistry (IHC) — Figure 2 Immunofluorescence characterization of transdifferentiated iCM (cardiac TF and microRNA). ( A ) Expression of ACTN2 and TNNT2 or ( B ) Expression of TNNT2 and Ki67 in HDF transdifferentiated for 2 weeks using induced expression of GATA4, TBX5, MEF2C, MYOCD, NKX2-5 and transfection with hsa-miR-1 and hsa-miR-133a. (Controls: M2rtTA only, microRNA only, cardiac TF only). ( C ) ACTN2. Panels on the right show varying degrees of cytoskeletal organization of ACTN2. ( D ) TNNT2. Panels on the right show varying degrees of cytoskeletal organization of TNNT2. ( E ) ACTN2 and GJA1. ( F ) TNNT2 and Ki67 expression in iCM. ( G ) VIM and MYH6/7. ( H ) TAGLN. ( I ) SMA. ( J ) MYH11. ( K ) ACTN2 expression in iCM 4 weeks following initiation of transdifferentiation. Panels on the right show varying degrees of cytoskeletal organization of ACTN2. ( L ) TNNT2 expression in iCM 4 weeks following initiation of transdifferentiation. Panels on the right show varying degrees of cytoskeletal organization of TNNT2.

Ki-67 Antibody in Western Blot (WB) — Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Western Blot

Ki-67 Antibody in Flow Cytometry (Flow) — Figure 1 PfRH5 vaccination with the ChAd63-MVA platform induces greater activation of total circulating CD8 + and CD4 + T cell populations, while the protein/AS01 B platform elicits a more sustained increase in cTfh-phenotype cells PBMCs from days 0, 7, 14, and 63 were stained ex vivo and analyzed using the gating strategies shown in Figures S1-S3 . (A and B) Increases at day 63 in activated CD38 + Ki67 + cells within total CD8 + and CD4 + T cells (A) or Th1 (CXCR3 + CCR6 - ), Th2 (CXCR3 - CCR6 - ), and Th17 (CXCR3 - CCR6 + ) CD4 + T cell subsets (B) were compared between platforms following subtraction of day 0 CD38 + Ki67 + frequencies in paired samples. (C-E) Frequencies of total cTfh cells defined as CXCR5 + (C) or CXCR5 + PD1 + (D) cells within the CD45RA - CD4 + T cell population were compared between platforms and the frequency of cTfr cells within total cTfh cells (defined as the CD25 + Foxp3 + subset) (E). All of the available samples are plotted (ChAd63-MVA/protein/AS01 B ): day 0, n = 15/54; day 7, n = 15/24; day 14, n = 15/54; day 63, n = 12/20. For intra-trial comparisons (E), only vaccinees with all 4 time points were analyzed: ChAd63-MVA, n = 12, protein/AS01 B , n = 17. Comparisons were performed with Mann-Whitney tests (between trials) or Friedman tests with Dunn's correction for multiple comparisons (within trials comparing day 0 to post-vaccination time points). *p < 0.5, **p < 0.01, and ***p < 0.001. In all of the panels, each point represents a vaccinee.

Ki-67 Antibody — 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 (20Raj1), eBioscience™ (Product # 14-5699-82) at 1 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), (1:2,000 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}

Product Details

14-5699-82

Applications	Tested Dilution	Publications
Immunohistochemistry (Paraffin) (IHC (P))	0.5-1 µg/mL	View 1 publication 1 publication
Immunocytochemistry (ICC/IF)	0.5-1 µg/mL	View 2 publications 2 publications
Flow Cytometry (Flow)	-	View 14 publications 14 publications
Immunohistochemistry (IHC)	-	View 3 publications 3 publications
Western Blot (WB)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Dog, Human
Published species	Human, Mouse
Host/Isotype	Mouse / IgG1, kappa
Class	Monoclonal
Type	Antibody
Clone	20Raj1
Conjugate	Unconjugated APC APC-eFluor 780 Biotin Brilliant UV 737 Brilliant UV 805 Brilliant Violet 605 eFluor 450 eFluor 506 eFluor 570 eFluor 615 eFluor 660 FITC PE PE-Cyanine5.5 PE-Cyanine7 PE-eFluor 610 PerCP-eFluor 710
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_2016711

Product Specific Information

Description: The monoclonal antibody 20Raj1 recognizes the human Ki-67 protein. Two isoforms of Ki-67 exist, a 345 and 395 kDa form that are expressed in dividing cells. Ki-67 is expressed in all cell types and is detectable during active phases of the cell cycle (G1, S, G2, and mitosis) but is absent from resting cells (G0). During interphase, Ki-67 expression is localized to the nucleus but redistributes to the chromosomes during mitosis and has specifically been found to associate with heterochromatin-bound proteins such as chromobox protein homolog 3 (CBX3). In studies of tumor cells, Ki-67 expression has been used as a marker for determining the fraction of proliferating cells within a given population of tumor cells.

This monoclonal antibody 20Raj1 recognizes canine (dog) Ki-67.

Applications Reported: This 20Raj1 antibody has been reported for use in immunocytochemistry and immunohistochemistry on formalin-fixed paraffin embedded tissues.

Applications Tested: This 20Raj1 antibody has been tested by immunocytochemistry on paraformaldehyde fixed and permeabilized MCF7, MDCK, and MDA-MB-175 cell lines.

This 20Raj1 antibody has been tested by immunohistochemistry on FFPE human skin using citrate buffer antigen retrieval. For immunocytochemistry and immunohistochemistry, this can be used at less than or equal to 0.5-1 µg/mL. It is recommended that the 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.

Product References

T cells drive negative feedback mechanisms in cancer associated fibroblasts, promoting expression of co-inhibitory ligands, CD73 and IL-27 in non-small cell lung cancer.

Oncoimmunology

O'Connor RA,Chauhan V,Mathieson L,Titmarsh H,Koppensteiner L,Young I,Tagliavini G,Dorward DA,Prost S,Dhaliwal K,Wallace WA,Akram AR

14-5699-82 was used in Flow Cytometry to study co-inhibitory ligands in T cell immunosuppression in non-small cell lung cancer.

Mon Aug 02 00:00:00 EDT 2021

Protein/AS01_B vaccination elicits stronger, more Th2-skewed antigen-specific human T follicular helper cell responses than heterologous viral vectors.

Cell reports. Medicine

Nielsen CM,Ogbe A,Pedroza-Pacheco I,Doeleman SE,Chen Y,Silk SE,Barrett JR,Elias SC,Miura K,Diouf A,Bardelli M,Dabbs RA,Barfod L,Long CA,Haynes BF,Payne RO,Minassian AM,Bradley T,Draper SJ,Borrow P

Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Flow Cytometry

Tue Mar 16 00:00:00 EDT 2021

Acetyl-CoA Synthetase 2: A Critical Linkage in Obesity-Induced Tumorigenesis in Myeloma.

Cell metabolism

Li Z,Liu H,He J,Wang Z,Yin Z,You G,Wang Z,Davis RE,Lin P,Bergsagel PL,Manasanch EE,Wong STC,Esnaola NF,Chang JC,Orlowski RZ,Yi Q,Yang J

14-5699-82 was used in Immunohistochemistry to demonstrate a key impact for obesity-induced acetyl-CoA synthetase 2 on the progression of myeloma.

Tue Jan 05 00:00:00 EST 2021

Single residue in CD28-costimulated CAR-T cells limits long-term persistence and antitumor durability.

The Journal of clinical investigation

Guedan S,Madar A,Casado-Medrano V,Shaw C,Wing A,Liu F,Young RM,June CH,Posey AD

Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Flow Cytometry

Mon Jun 01 00:00:00 EDT 2020

Functional Study of the Retrotransposon-Derived Human PEG10 Protease.

International journal of molecular sciences

Golda M,Mótyán JA,Mahdi M,Tőzsér J

Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Flow Cytometry

Tue Mar 31 00:00:00 EDT 2020

Human Liver Memory CD8⁺ T Cells Use Autophagy for Tissue Residence.

Cell reports

Swadling L,Pallett LJ,Diniz MO,Baker JM,Amin OE,Stegmann KA,Burton AR,Schmidt NM,Jeffery-Smith A,Zakeri N,Suveizdyte K,Froghi F,Fusai G,Rosenberg WM,Davidson BR,Schurich A,Simon AK,Maini MK

Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Flow Cytometry

Tue Jan 21 00:00:00 EST 2020

Circulating Tumor Cells Exhibit Metastatic Tropism and Reveal Brain Metastasis Drivers.

Cancer discovery

Klotz R,Thomas A,Teng T,Han SM,Iriondo O,Li L,Restrepo-Vassalli S,Wang A,Izadian N,MacKay M,Moon BS,Liu KJ,Ganesan SK,Lee G,Kang DS,Walmsley CS,Pinto C,Press MF,Lu W,Lu J,Juric D,Bardia A,Hicks J,Salhia B,Attenello F,Smith AD,Yu M

14-5699-82 was used in Immunohistochemistry (Paraffin) to demonstrate that ex vivo-cultured CTCs from 4 patients with breast cancer showed organotropism, revealing molecular features that allow a subset of CTCs to enter and grow in the brain.

Wed Jan 01 00:00:00 EST 2020

Niraparib activates interferon signaling and potentiates anti-PD-1 antibody efficacy in tumor models.

Scientific reports

Wang Z,Sun K,Xiao Y,Feng B,Mikule K,Ma X,Feng N,Vellano CP,Federico L,Marszalek JR,Mills GB,Hanke J,Ramaswamy S,Wang J

Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Flow Cytometry

Tue Feb 12 00:00:00 EST 2019

Streptomyces Levis ABRIINW111 Inhibits SW480 Cells Growth by Apoptosis Induction.

Advanced pharmaceutical bulletin

Faramarzian Azimi Maragheh B,Fatourachi P,Mohammadi SM,Valipour B,Behtari M,Dehnad A,Nozad Charoudeh H

Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Flow Cytometry

Thu Nov 01 00:00:00 EDT 2018

Tissue-resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer.

Clinical and experimental immunology

Hartana CA,Ahlén Bergman E,Broomé A,Berglund S,Johansson M,Alamdari F,Jakubczyk T,Huge Y,Aljabery F,Palmqvist K,Holmström B,Glise H,Riklund K,Sherif A,Winqvist O

14-5699 was used in Flow cytometry/Cell sorting to identify TRM cells as potential new targets for cancer immunotherapy.

Mon Oct 01 00:00:00 EDT 2018

Highly multiplexed immunofluorescence imaging of human tissues and tumors using t-CyCIF and conventional optical microscopes.

eLife

Lin JR,Izar B,Wang S,Yapp C,Mei S,Shah PM,Santagata S,Sorger PK

Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Flow Cytometry

Wed Jul 11 00:00:00 EDT 2018

Involvement of TWEAK and the NF-κB signaling pathway in lupus nephritis.

Experimental and therapeutic medicine

Sun F,Teng J,Yu P,Li W,Chang J,Xu H

Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Flow Cytometry

Thu Mar 01 00:00:00 EST 2018

Small-Vessel Vasculopathy Due to Aberrant Autophagy in LAMP-2 Deficiency.

Scientific reports

Nguyen HT,Noguchi S,Sugie K,Matsuo Y,Nguyen CTH,Koito H,Shiojima I,Nishino I,Tsukaguchi H

Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Flow Cytometry

Tue Feb 20 00:00:00 EST 2018

Core Transcription Factors, MicroRNAs, and Small Molecules Drive Transdifferentiation of Human Fibroblasts Towards The Cardiac Cell Lineage.

Scientific reports

Christoforou N,Chakraborty S,Kirkton RD,Adler AF,Addis RC,Leong KW

14569982 was used in immunocytochemistry to describe the transdifferentiation of human dermal fibroblasts towards the cardiac cell lineage

Tue Jan 10 00:00:00 EST 2017

Core Transcription Factors, MicroRNAs, and Small Molecules Drive Transdifferentiation of Human Fibroblasts Towards The Cardiac Cell Lineage.

Scientific reports

Christoforou N,Chakraborty S,Kirkton RD,Adler AF,Addis RC,Leong KW

14569982 was used in immunocytochemistry to describe the transdifferentiation of human dermal fibroblasts towards the cardiac cell lineage

Tue Jan 10 00:00:00 EST 2017

S100A6 Regulates Endothelial Cell Cycle Progression by Attenuating Antiproliferative Signal Transducers and Activators of Transcription 1 Signaling.

Arteriosclerosis, thrombosis, and vascular biology

Lerchenmüller C,Heißenberg J,Damilano F,Bezzeridis VJ,Krämer I,Bochaton-Piallat ML,Hirschberg K,Busch M,Katus HA,Peppel K,Rosenzweig A,Busch H,Boerries M,Most P

Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Immunocytochemistry

Thu Sep 01 00:00:00 EDT 2016

S100A6 Regulates Endothelial Cell Cycle Progression by Attenuating Antiproliferative Signal Transducers and Activators of Transcription 1 Signaling.

Arteriosclerosis, thrombosis, and vascular biology

Lerchenmüller C,Heißenberg J,Damilano F,Bezzeridis VJ,Krämer I,Bochaton-Piallat ML,Hirschberg K,Busch M,Katus HA,Peppel K,Rosenzweig A,Busch H,Boerries M,Most P

Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Immunocytochemistry

Thu Sep 01 00:00:00 EDT 2016

Interleukin-7 and Toll-like receptor 7 induce synergistic B cell and T cell activation.

PloS one

Bikker A,Kruize AA,van der Wurff-Jacobs KM,Peters RP,Kleinjan M,Redegeld F,de Jager W,Lafeber FP,van Roon JA

Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Flow Cytometry

Sat Feb 07 00:00:00 EST 2015

Biliary tree stem cells, precursors to pancreatic committed progenitors: evidence for possible life-long pancreatic organogenesis.

Stem cells (Dayton, Ohio)

Wang Y,Lanzoni G,Carpino G,Cui CB,Dominguez-Bendala J,Wauthier E,Cardinale V,Oikawa T,Pileggi A,Gerber D,Furth ME,Alvaro D,Gaudio E,Inverardi L,Reid LM

Published figure using Ki-67 monoclonal antibody (Product # 14-5699-82) in Immunofluorescence

Sun Sep 01 00:00:00 EDT 2013

Somatostatin expression in human hair follicles and its potential role in immune privilege.

The Journal of investigative dermatology

Breitkopf T,Lo BK,Leung G,Wang E,Yu M,Carr N,Zloty D,Cowan B,Shapiro J,McElwee KJ

14-5699 was used in Flow cytometry/Cell sorting to demonstrate and quantify the functional immune privilege properties apparent in human hair follicles.

Mon Jul 01 00:00:00 EDT 2013

Dll4-Notch signaling in Flt3-independent dendritic cell development and autoimmunity in mice.

The Journal of experimental medicine

Billiard F,Lobry C,Darrasse-Jèze G,Waite J,Liu X,Mouquet H,DaNave A,Tait M,Idoyaga J,Leboeuf M,Kyratsous CA,Burton J,Kalter J,Klinakis A,Zhang W,Thurston G,Merad M,Steinman RM,Murphy AJ,Yancopoulos GD,Aifantis I,Skokos D

14-5699 was used in Flow cytometry/Cell sorting to investigate how Dll4-Notch signaling affects pro-T cell fate and thymic DC development, showing that it is a novel Flt3-independent pathway for DC development and autoimmunity.

Mon May 07 00:00:00 EDT 2012

Bioinformatics

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

Gene Aliases: KIA; MIB-; MIB-1; MKI67; PPP1R105

UniProt ID: (Human) P46013

Entrez Gene ID: (Dog) 100686578, (Human) 4288

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

Ki-67 Monoclonal Antibody (20Raj1), eBioscience™

Product Details

14-5699-82

Immunohistochemistry (Paraffin) (IHC (P))

Immunocytochemistry (ICC/IF)

Flow Cytometry (Flow)

Immunohistochemistry (IHC)

Western Blot (WB)

Species Reactivity

Published species

Host/Isotype

Class

Type

Clone

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

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