Ki67 16a8

Manufactured by BioLegend

Ki67 (16A8) is a monoclonal antibody that recognizes the Ki-67 protein, a cellular marker for proliferation. Ki-67 is a nuclear protein associated with and required for cellular proliferation.

Automatically generated - may contain errors

Lab products found in correlation

Foxp3 fjk 16s, by Thermo Fisher Scientific (3 mentions) Cd62l mel 14, by BioLegend (2 mentions) Cytofix cytoperm fixation permeabilization kit, by BD (1 mentions) Cd11b m1 70, by BioLegend (1 mentions) Anti cd3 145 2c11, by Thermo Fisher Scientific (1 mentions) Anti cd28 37.51, by Thermo Fisher Scientific (1 mentions) Trail n2b2, by Thermo Fisher Scientific (1 mentions) Cd8 53 6, by Thermo Fisher Scientific (1 mentions) Klrg1 2f1 klrg1, by BioLegend (1 mentions) Cd43 1b11, by BioLegend (1 mentions) Cd4 rm4 5, by BioLegend (1 mentions) Fixable viability dye, by Thermo Fisher Scientific (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Foxp3 transcription factor staining kit, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions) Facs lsr 2, by BD (1 mentions) Mouse regulatory t cell staining kit, by Thermo Fisher Scientific (1 mentions) Ctla 4 uc10 4b9, by BioLegend (1 mentions) Helios 22f6, by BioLegend (1 mentions) Lsrfortessa, by BD (1 mentions)

8 protocols using ki67 16a8

Comprehensive Immune Cell Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell surface and intracellular stainings were performed using antibodies as follows: CD4 (RM 4–5; BioLegend), CD8 (53–6.7; eBioscience), CD43 (1B11; BioLegend), CD62L (MEL-14; BioLegend), CD11b (M1/70; BioLegend), KLRG1 (2F1/KLRG1; BioLegend), Ki67 (16A8; BioLegend), FasL (MFL3; BD Biosciences), TRAIL (N2B2; eBioscience), Foxp3 (FJK-16s; eBioscience), GzmB (GB12; ThermoFisher Scientific), GzmA (GzA-368.5; eBioscience) and Gzm K (Orb102688; Biorbyt). Dead cells were excluded by fixable viability dye (eBioscience) staining. For FasL staining, lymphocytes were isolated and restimulated with anti-CD3 (145-2C11, eBioscience) and anti-CD28 (37.51, eBioscience) antibodies for 5 hours at 37 °C. BD Cytofix/Cytoperm Fixation/Permeabilization kit was used for intracellular staining following the manufacturer’s instructions. Data were acquired on an LSR II flow cytometer (BD Biosciences). Analyses were done using FlowJo 5.0 software (Tree Star Inc., Ashland, OR).

Malyshkina A., Littwitz-Salomon E., Sutter K., Zelinskyy G., Windmann S., Schimmer S., Paschen A., Streeck H., Hasenkrug K.J, & Dittmer U. (2017). Fas Ligand-mediated cytotoxicity of CD4+ T cells during chronic retrovirus infection. Scientific Reports, 7, 7785.

+ Open protocol

+ Expand

Comprehensive Immunophenotyping Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following clones of antibody were purchased from Biolegend (San Diego, CA, USA) and used for cell surface staining: CD4 (RM4-5), TCRβ (H57-597), NK1.1 (PK136), CD44 (IM7), CD24 (M1/69), CD127 (A7R34), CD69 (H1.2F3), CD25 (PC61), CD5 (53-7.3). For nuclear protein staining, cells were fixated and permeabilized using the FOXP3 transcription factor staining kit (Thermo Fisher Scientific, Waltham, MA, USA), according to the manufacturer’s instructions. The following antibodies were used: Ki67 (16A8) and PLZF (Mags.21F7) were purchased from Biolegend. Flow cytometry was performed on a FACSCanto II or FACSLSRII (BD Bioscience, San Jose, CA, USA) and analyzed with FlowJo software (version 10.8.1, BD Bioscience, San Jose, CA, USA). CD1d tetramer, either loaded or unloaded with αGC, was obtained from NIH Tetramer Core Facility (Atlanta, GA, USA).

Chuang Y.T., Chuang W.C., Liu C.C., Liu C.W., Huang Y.W., Yang H.Y., Ho I.C, & Tai T.S. (2021). Ets1 Promotes the Differentiation of Post-Selected iNKT Cells through Regulation of the Expression of Vα14Jα18 T Cell Receptor and PLZF. International Journal of Molecular Sciences, 22(22), 12199.

+ Open protocol

+ Expand

Comprehensive Treg Immunophenotyping Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Dead cells were stained using the fixable viability dye ZombieRed according to manufacturer’s instructions. Chemokine receptors CCR4 (clone 2G12), CXCR3 (CXCR3–173) and CCR5 (HM-CCR5) were stained for 1 hour at 37°C in cell culture medium. Staining for CD4 (GK1.5d), CD25 (PC61), CD44 (IM7), CD45 (30F11), CD62L (MEL14), ICOS (C398.4A), was performed at 4°C for 20 min. All these antibodies were from Biolegend. To detect intracytoplasmic and intranuclear antigens, cells were fixed and permeabilized using the Mouse Regulatory T cell Staining Kit from eBioscience, and stained for Ki67 (16A8, Biolegend), CTLA-4 (UC10-4B9, Biolegend), GFP/YFP (Alexa Fluor 488-conjugated polyclonal Rabbit IgG, Life Technologies), and Foxp3 (FJK-16s, eBioscience).
For some experiments, as indicated, blood-borne were distinguished from tissue-resident Treg by an i.v. injection of 3µg CD45.2-PE mAb (clone 104) 3 minutes before euthanasia.

Marangoni F., Zhang R., Mani V., Thelen M., Ali Akbar N.J., Warner R.D., Äijö T., Zappulli V., Martinez G.J., Turka L.A, & Mempel T.R. (2018). Tumor tolerance-promoting function of regulatory T cells is optimized by CD28, but strictly dependent on calcineurin. Journal of immunology (Baltimore, Md. : 1950), 200(10), 3647-3661.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry for Immune Cell Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Samples were analyzed using Aurora (Cytek) and LSR Fortessa (Becton Dickinson) flow cytometers and the resulting data were analyzed using FlowJo software (Tree Star, Inc). The following monoclonal antibodies were used from eBioscience, CD3ε (145-2C11); CD4 (GK1.5); CD8a (53-6.7); CD45 (30-F11); Foxp3 (FJK-16s). Helios (22F6) and Ki67 (16A8) were from Biolegend. GITR was from BD. Nrp1 was purchased from R&D Systems.

Centa M., Weinstein E.G., Clemente J.C., Faith J.J., Fiel M.I., Lyallpuri R., Herbin O, & Alexandropoulos K. (2022). Impaired Central Tolerance Induces Changes in the Gut Microbiota that Exacerbate Autoimmune Hepatitis. Journal of autoimmunity, 128, 102808.

+ Open protocol

+ Expand

Comprehensive Immune Cell Phenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tissues were processed, single cell suspensions obtained, and cells were stained as described (Wherry et al., 2003). Cells were stained with LIVE/DEAD cell stain (Invitrogen) and with antibodies targeting KLRG1 (2F1/KLRG1, Biolegend), CD127 (A7R34, Biolegend), CD8 (53-6.7, Biolegend), CD44 (IM7, Biolegend), CD45.1 (A20, Biolegend), CD45.2 (104, Biolegend), CD62L (MEL-14, Biolegend), CD27 (LG.7F9, Biolegend), CD122 (TM-b1, eBioscience), CXCR3 (CXCR3-173, Biolegend), Bcl-2 (A19-3, BD PharMingen), Bcl-xL (H-5, Santa Cruz), Bcl-6 (K112-91, BD PharMingen), Eomes (Dan11mag, eBioscience), Ki-67 (16A8, BioLegend), Bim (C34C5, Cell Signaling Transduction), and MHC class I D^bgp^33–41 tetramer (NIH tetramer core). Intracellular cytokine staining was performed after 5 hrs ex vivo stimulation with gp^33–41 peptide in the presence of GolgiPlug (BD), GolgiStop (BD) and CD107a (1D4B, Biolegend). After stimulation, cells were stained with surface antibodies, following fixation with Fixation/Permeabilization Buffer (eBioscience) and then stained with intracellular antibodies for TNF (MP6-XT22, Biolegend), IFN-γ (XMG1.2, BD PharMingen), GrzmB (GRB17, Life Technologies) and MIP-1α (IC450P, R&D System) using Permeabilization Wash Buffer (eBioscience) according to manufacturer’s instructions. Cells were analyzed with LSRII (BD Biosciences) and FlowJo software (Treestar).

Chen Z., Stelekati E., Kurachi M., Yu S., Cai Z., Manne S., Khan O., Yang X, & Wherry E.J. (2017). MiR-150 regulates memory CD8 T cell differentiation via c-Myb. Cell reports, 20(11), 2584-2597.

+ Open protocol

+ Expand

Multi-parameter Flow Cytometric Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Single-cell suspensions from the indicated organs were treated with anti-CD16/32 (clone 93; Biolegend, San Diego, CA, USA) for Fc-blocking. The following antibodies were used for cell-surface staining: anti-B220 (RA3-6B2), CD19 (6D5); IgM (RMM-1); IgD (11-26c.2a); CD5 (53-7.3); CD21 (7E9); CD23 (B3B4), CD22 (OX-97); PIR-A/B (6C1); BAFFR (7H22-E16); CD3ε (145-2c11); CD4 (GK1.5); CD8α (53-6.7); CD11b (M1/70); Gr-1 (RB6-8c5); Ter-119 (TER-119); CD95 (Jo2); GL7 (GL7); CD138 (281-2); CD38 (90); IgG₁ (RMG1-1); and Ki67 (16A8) (all from Biolegend). Anti-phospho-Btk (pY223: N35-86) and PLCγ2 (pY759: K86-689.37) antibodies were from BD Biosciences (San Jose, CA, USA). Debris and dead cells were excluded by forward scatter, side scatter and PI staining. All samples were analyzed by FACSCalibur or FACSAria (BD Biosciences). The data were analyzed using FlowJo (TreeStar, Ashland, OR, USA).

Sasanuma H., Ozawa M, & Yoshida N. (2018). RNA-binding protein Ptbp1 is essential for BCR-mediated antibody production. International Immunology, 31(3), 157-166.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry Analysis of Immune Cell Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse spleen and PLN were mechanically dissociated into single cells with use of 40-μm filters, and red cell lysis was performed with ACK buffer. Blocking was performed with anti-CD16/CD32 antibodies (BD Biosciences), and cells were stained with antibodies against CD4 (RM4-5; BD Biosciences), CD8 (53-6.7; BD Biosciences), CD11b (M1/70; BD Biosciences), CD11c (HL3; BD Biosciences), B220 (RA3-6B2; BD Biosciences), F4/80 (CI:A3-1; Bio-Rad Laboratories), CD62L (MEL-14; BD Biosciences), CD44 (IM7; BD Biosciences), CD25 (PC61; BD Biosciences), FoxP3 (FJK-16s; eBioscience), TIGIT (1G9; BioLegend), KLRG1 (2F1; BioLegend) and Ki67 (16A8; BioLegend). Samples were analyzed on the LSRII (BD Biosciences) and FlowJo (Tree Star, Inc.).

Leung S.S., Borg D.J., McCarthy D.A., Boursalian T.E., Cracraft J., Zhuang A., Fotheringham A.K., Flemming N., Watkins T., Miles J.J., Groop P.H., Scheijen J.L., Schalkwijk C.G., Steptoe R.J., Radford K.J., Knip M, & Forbes J.M. (2022). Soluble RAGE Prevents Type 1 Diabetes Expanding Functional Regulatory T Cells. Diabetes, 71(9), 1994-2008.

+ Open protocol

+ Expand

Oncolytic Adenovirus and Anti-PD-1 Therapy

Check if the same lab product or an alternative is used in the 5 most similar protocols

Oncolytic adenovirus (OV) is a type 5 adenovirus with depleted E1b and E3 genes and a retained E1a gene. The human telomerase reverse transcriptase promoter was inserted upstream of the E1A gene to control virus replication. The OV was packaged and amplified in 293T cells, and ultracentrifugation was used for purification. Therapeutic anti-PD-1 antibodies were provided by Innovent Biologics (Suzhou, Jiangsu, China). Fluorescence-labeling antibodies to CD3 (17A2), CD4 (RM4-5), CD45 (30-F11), CD11b (M1/70), F4/80 (BM8), CD206 (C068C2), LAG-3 (C9B7W), PD-1 (RMP1-30), ICOS (7E.17G9), CD44 (IM7), CD62L (MEL-14), MHCII (M5/114.15.2), and Ki67 (16A8) were purchased from BioLegend, and antibodies to CD8 (53-6.7), FoxP3 (MF23), and IFN-γ (XMG1.2) were purchased from BD Bioscience. Antibody to adenovirus-5 E1A was purchased from Santa Cruz Biotechnology. OVA Tetramer-SIINFEKL-PE and fluorescein isothiocyanate-labeling anti-CD8 (KT15) were purchased from MBL.

Shi G., Shi P., Yu Y., Xu J., Ma J., Zhang Y., Dong Z., Shen L., Dai L., Cheng L., Cheng P, & Deng H. (2021). Oncolytic adenovirus inhibits malignant ascites of advanced ovarian cancer by reprogramming the ascitic immune microenvironment. Molecular Therapy Oncolytics, 23, 488-500.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!