Cytoperm solution

Manufactured by BD

BD Cytoperm solution is a permeabilization reagent used in flow cytometry applications to allow intracellular staining of cells. It facilitates the entry of antibodies or other probes into the cell interior for the detection of intracellular proteins or other cellular components.

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Lab products found in correlation

Chamber slide, by Thermo Fisher Scientific (1 mentions) Lsm780 confocal laser scanning microscope, by Zeiss (1 mentions) Cell staining buffer, by BioLegend (1 mentions) Apc anti human cd69, by BioLegend (1 mentions) Live dead fixable yellow dead cell stain, by Thermo Fisher Scientific (1 mentions) Apc cy7 anti human cd3, by BioLegend (1 mentions) Facscelesta flow cytometer, by BD (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Cytofix, by BD (1 mentions)

2 protocols using cytoperm solution

Immunofluorescence analysis of CSDE1 in B16 cells

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Uninfected B16 parental and B16 parental cells infected with VSV-IFN-β (MOI = 0.1) for 8 h, then seeded in chamber slides (Thermo Fisher Scientific, cat. no. 154534) at 20,000 cells per well. Cells were allowed to attach overnight, then medium was removed, cell monolayer washed once with PBS, then fixed for 20 min (BD Biosciences, cat. no. 555028). Cells were then permeabilized and blocked for 1 h using BD Cytoperm solution. Polyclonal anti-CSDE1 antibody (Atlas Antibodies, cat. no. HPA052221) treatment was done overnight (∼14 h) in humidity chambers to prevent drying. Slides were washed three times with PBS and then incubated with species-appropriate Alexa Fluor 594-IgG (H + L) (Jackson ImmunoResearch, cat. no. 711-586-152) for 60 min in the presence of DAPI. Cells were then observed with a Zeiss LSM 780 confocal laser scanning microscope and the images were analyzed with Zeiss imaging software.

Webb M.J., Kottke T., Kendall B.L., Swanson J., Uzendu C., Tonne J., Thompson J., Metko M., Moore M., Borad M., Roberts L., Diaz R.M., Olin M., Borgatti A, & Vile R. (2023). Trap and ambush therapy using sequential primary and tumor escape-selective oncolytic viruses. Molecular Therapy Oncolytics, 29, 129-142.

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PBMC Isolation and Characterization from Colon Chip

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PBMC were harvested from Colon Intestine-Chip epithelial channels at the final timepoint after administration. PBMC were washed with DPBS and stained with 2 μM live/dead Fixable Yellow Dead Cell Stain (ThermoFisher) and washed in DPBS. PBMC were then fixed with BD Cytofix (BD Biosciences) fixation solution, washed in DPBS, and either resuspended in 90% FBS (Sigma) + 10% DMSO solution and frozen at −20°C until use or stained immediately. Samples to be stained for surface markers were washed in DPBS and resuspended in Cell Staining Buffer (Biolegend). Surface marker stains were prepared in BD /Cytoperm solution (BD Biosciences) and consisted of anti-human CD3 APC-Cy7 (BioLegend), anti-human CD4 Brilliant Violet 786 (BioLegend), anti-human-CD8-PE/Dazzle-594 (BioLegend), and anti-human CD69 APC (BioLegend).
Sample data was acquired using the BD FACSCelesta flow cytometer (BD Biosciences), and data was analyzed using FlowJo V10 software (FlowJo).

Kerns S.J., Belgur C., Petropolis D., Kanellias M., Barrile R., Sam J., Weinzierl T., Fauti T., Freimoser-Grundschober A., Eckmann J., Hage C., Geiger M., Ng P.R., Tien-Street W., Manatakis D.V., Micallef V., Gerard R., Bscheider M., Breous-Nystrom E., Schneider A., Giusti A.M., Bertinetti-Lapatki C., Grant H.S., Roth A.B., Hamilton G.A., Singer T., Karalis K., Moisan A., Bruenker P., Klein C., Bacac M., Gjorevski N, & Cabon L. (2021). Human immunocompetent Organ-on-Chip platforms allow safety profiling of tumor-targeted T-cell bispecific antibodies. eLife, 10, e67106.

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