Perforin fitc

Manufactured by Miltenyi Biotec

Perforin-FITC is a fluorochrome-conjugated antibody used for the detection and analysis of perforin, a cytotoxic protein involved in the killing of target cells. The FITC (fluorescein isothiocyanate) label allows for the visualization and quantification of perforin-expressing cells through flow cytometry or other fluorescence-based techniques.

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Lab products found in correlation

Facsverse cytometer, by BD (1 mentions) Granzyme b pe, by Miltenyi Biotec (1 mentions) Hla dr vioblue, by Miltenyi Biotec (1 mentions) Cd38 pevio770, by Miltenyi Biotec (1 mentions) Cd4 apc vio770, by Miltenyi Biotec (1 mentions) Cd8 apc, by Miltenyi Biotec (1 mentions) Epoch microplate spectrophotometer, by Agilent Technologies (1 mentions) Human male ab serum, by Merck Group (1 mentions) Stem cell growth medium, by CellGenix (1 mentions) Cytofix cytoperm kit, by BD (1 mentions)

2 protocols using perforin fitc

Phenotyping of Activated PBMC Subsets

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1.5 × 10^{^6} of thawed PBMCs were plated in complete RPMI containing 10% human serum supplemented with 1% Penicillin–Streptomycin–Glutamin. Overnight-rested PBMCs were stained with the appropriate antibodies for 20 min at 4 °C in the dark and acquired using FACSVerse™cytometer (BD Biosciences). Dead cells were labeled using ViobilityTM Fixable Dye (Miltenyi Biotec). Antibodies used were: CD4-APC-Vio770, CD8-APC, HLA-DR-VioBlue, CD38-PE-Vio770, Granzyme B-PE and Perforin-FITC (Miltenyi Biotec). (Representative plots are shown in Supplementary Fig. 6). Data were analyzed using FlowJo 10.7.2 (BD Biosciences).

Rovito R., Bono V., Augello M., Tincati C., Mainoldi F., Beaudoin-Bussières G., Tauzin A., Bianchi S., Hadla M., Yellenki V., d’Arminio Monforte A., Casola S., Borghi E., Finzi A, & Marchetti G. (2022). Association between SARS-CoV-2 RNAemia and dysregulated immune response in acutely ill hospitalized COVID-19 patients. Scientific Reports, 12, 19658.

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NK Cell-Mediated Cytotoxicity Assay

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K562mbIL15-41BBL feeder cell line (10⁷ cells) was co-cultured with 1.5 × 10⁷ PBMCs for 2 weeks in stem cell growth medium (Cellgenix), with 10% human male AB serum (Sigma) to obtain highly purified NK cells. IL-2 (10 IU/mL) was added for the first week and 100 IU/mL thereafter. NK cells were then exposed to both sorted GFP⁺ and CD14⁺GFP⁺ cellular entities. After cell permeabilization (Cytofix/Cytoperm kit, BD Biosciences), CD3⁻CD56⁺CD16⁺ NKs perforin generation was detected with specific Perforin-FITC (Miltenyi-130 096 668). Cytotoxicity was monitored with conventional 2 h Europium lysis-terpyridine dicarboxylic acid (TDA) release assay (Perkin-Elmer). NK (effector) cells loaded TDA-labeled (target) H460-CSC-GFP⁺ and CD14⁺GFP⁺ hybrid cells, at effector-to-target cell ratios of 50:1, 25:1, 12.5:1 and 6.25:1. Fluorescent measures (ex: 340 nm; em: 612 nm) were assessed in a BioTek Epoch Microplate Spectrophotometer.

Aguirre L.A., Montalbán-Hernández K., Avendaño-Ortiz J., Marín E., Lozano R., Toledano V., Sánchez-Maroto L., Terrón V., Valentín J., Pulido E., Casalvilla J.C., Rubio C., Diekhorst L., Laso-García F., del Fresno C., Collazo-Lorduy A., Jiménez-Munarriz B., Gómez-Campelo P., Llanos-González E., Fernández-Velasco M., Rodríguez-Antolín C., Pérez de Diego R., Cantero-Cid R., Hernádez-Jimenez E., Álvarez E., Rosas R., dies López-Ayllón B., de Castro J., Wculek S.K., Cubillos-Zapata C., Ibáñez de Cáceres I., Díaz-Agero P., Gutiérrez Fernández M., Paz de Miguel M., Sancho D., Schulte L., Perona R., Belda-Iniesta C., Boscá L, & López-Collazo E. (2020). Tumor stem cells fuse with monocytes to form highly invasive tumor-hybrid cells. Oncoimmunology, 9(1), 1773204.

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