Perm2 permeabilizing solution

Manufactured by BD

BD Perm2 Permeabilizing Solution is a laboratory reagent used to temporarily permeabilize cell membranes, allowing for the intracellular staining and analysis of cellular components. It is a key component in various cell analysis techniques.

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Lab products found in correlation

Facs lysing solution, by BD (2 mentions) Dead cell marker, by Thermo Fisher Scientific (2 mentions) Lsrfortessa, by BD (1 mentions)

Spelling variants of this product

Perm2 (12 citations) Permeabilizing solution (10 citations)

2 protocols using perm2 permeabilizing solution

Multiparameter Flow Cytometric Analysis

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Freshly isolated PBMCs were stained with dead cell marker (Invitrogen, Carlsbad) and fluorochrome‐conjugated antibodies directed against intracellular and surface markers (Supplementary table 1) for 20 min at RT in the dark. Cells were washed two times with 150 µL of PBS with 10% FCS. Next, pellets were resuspended in 100 µL of BD FACS Lysing Solution (BD Biosciences) and incubated for 10 min at RT, for cell fixation. After washing with PBS with 10% FCS, cells were permeabilised using 100 µL of BD Perm2 Permeabilizing Solution (BD Biosciences) for 10 min at RT. Subsequently, antibodies were added for intracellular staining and incubated for 30 min at RT in the dark. Cells were then washed with 150 µL of PBS with 10% FCS and incubated in a 1% formaldehyde solution for 2 h, washed and resuspended in PBS containing 10% FCS.
Samples were acquired on a BD LSR Fortessa™. Flow cytometric analysis was performed using FlowJo version 10.6.2 (TreeStar, Ashland).

García M., Kokkinou E., Carrasco García A., Parrot T., Palma Medina L.M., Maleki K.T., Christ W., Varnaitė R., Filipovic I., Ljunggren H., Björkström N.K., Folkesson E., Rooyackers O., Eriksson L.I., Sönnerborg A., Aleman S., Strålin K., Gredmark‐Russ S., Klingström J, & Mjösberg J. (2020). Innate lymphoid cell composition associates with COVID‐19 disease severity. Clinical & Translational Immunology, 9(12), e1224.

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Detailed PBMC Immunophenotyping Protocol

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Freshly isolated PBMCs were stained with dead cell marker (Invitrogen) and fluorochrome-conjugated antibodies directed against intracellular and surface markers (See Table S1 in the Online Repository) for 20 min at RT in the dark. Cells were washed two times with 150 ul of PBS with 10% FCS. Next, pellets were resuspended in 100 ul of BD FACS Lysing Solution (BD Biosciences) and incubated for 10 min at RT, for cell fixation. After washing with PBS with 10% FCS, cells were permeabilized using 100 ul of BD Perm2 Permeabilizing Solution (BD Biosciences) for 10 min at RT. Subsequently, antibodies were added for intracellular staining and incubated for 30 min at RT in the dark. Cells were then washed with 150 ul of PBS with 10% FCS and incubated in a 1% formaldehyde solution for 2h, washed and resuspended in PBS containing 10% FCS.
The antibodies and dead cell marker used for flow cytometry staining are listed in Supplementary Table III. Samples were acquired on a BD LSR Fortessa TM . Flow cytometric analysis was performed using FlowJo version 10.6.2 (TreeStar).

García M., Kokkinou E., García A.C., Parrot T., Medina L.M.P., Maleki K.T., Christ W., Varnaitė R., Filipovic I., Ljunggren H., Björkström N.K., Folkesson E., Rooyackers O., Eriksson L.I., Sönnerborg A., Aleman S., Strålin K., Gredmark‐Russ S., Klingström J., & Mjösberg J. (2020). Innate lymphoid cell composition associates with COVID-19 disease severity.

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