Permeabilization buffer 10x

Manufactured by Thermo Fisher Scientific

Sourced in United States

Permeabilization Buffer 10X is a laboratory reagent designed to temporarily permeabilize cell membranes. It is a concentrated solution that is diluted to 1X working concentration prior to use. The buffer facilitates the delivery of molecules, such as antibodies or nucleic acids, into the interior of cells for analysis or other applications.

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Lab products found in correlation

Facsdiva software, by BD (1 mentions) Lrsfortessa flow cytometer, by BD (1 mentions) Live dead fixable aqua dead cell stain, by Thermo Fisher Scientific (1 mentions) Fixation perm buffer, by Thermo Fisher Scientific (1 mentions) Pe ido, by Thermo Fisher Scientific (1 mentions) Cd8 apc h7, by BD (1 mentions) Cd127 percp cy5, by BD (1 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) Cd44 pe, by BD (1 mentions) Cd45 v500, by BD (1 mentions) Foxp3 fitc, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions) Ionomycin, by Merck Group (1 mentions) Red blood cell lysis buffer, by BioLegend (1 mentions) Trypan blue, by Merck Group (1 mentions) Fetal bovine serum (fbs), by PAN Biotech (1 mentions) Cd62l apc, by BD (1 mentions) Brefeldin a, by BD (1 mentions) Cd69 apc, by BD (1 mentions)

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Permeabilization buffer (703 citations)

3 protocols using permeabilization buffer 10x

Multiparametric Flow Cytometry for DC Subsets

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For DCs flow cytometry, PBMCs were centrifuged, pelleted and washed with Phosphate-buffered saline (PBS) and stained for 35 min at room temperature with LIVE/DEAD Fixable Aqua Dead Cell Stain (Life Technologies), BV421 CD86, BV650 CD11c, BV711 HLA-DR, BV786 CCR7 (CD197), FITC Lin-2 (CD3, CD14, CD19, CD20, CD56), BV605 CD16, PeCF594 PD-L1 (CD274), APC Integrin-β7 (BD Biosciences), PerCPCy5,5 CD4, APCCy7 CD1c, PeCy7 CD141 (BioLegend) and AF700 CD123 (R&D, San Diego, CA) antibodies. Then PBMCs were washed with Permeabilization Buffer 10X diluted 1:10 (eBioscience™), permeabilized by Fixation/Perm buffer (eBioscience™), and intracellularly stained with PE IDO (eBioscience, San Diego, CA, USA) antibody. DCs were gated based on Lin-2 HLA-DR expression. Each subset (mDCs and pDCS) was gated based on CD123 and CD11c expression. mDCs subsets were gated by using CD16, CD1c and CD141 staining, for gating strategy see Supplementary Fig. 1. Flow cytometry analyses were performed on an LRS Fortessa flow cytometer using FACS Diva software (BD Biosciences). Data were analyzed using the FlowJo software (Treestar, Ashland, OR). At least 1 × 10⁶ events were acquired per sample.

Pérez-Gómez A., Vitallé J., Gasca-Capote C., Gutierrez-Valencia A., Trujillo-Rodriguez M., Serna-Gallego A., Muñoz-Muela E., Jiménez-Leon M.D., Rafii-El-Idrissi Benhnia M., Rivas-Jeremias I., Sotomayor C., Roca-Oporto C., Espinosa N., Infante-Domínguez C., Crespo-Rivas J.C., Fernández-Villar A., Pérez-González A., López-Cortés L.F., Poveda E, & Ruiz-Mateos E. (2021). Dendritic cell deficiencies persist seven months after SARS-CoV-2 infection. Cellular and Molecular Immunology, 18(9), 2128-2139.

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Flow Cytometry Analysis of Mouse and Human T Cells

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Anti-human CD3e-V500, CD4-APC-H7, CD8-APC-H7, CD25-PE, CD39-PE (clone TU66), CD73-APC (clone AD2), PD1-PE-Cy7 (clone EH12.1), and CD127-PerCP-Cy5.5 antibodies were purchased by BD Biosciences (Germany), anti-human FOXP3-FITC antibody was from eBioscience (Germany). Anti-mouse CD3e-PerCP-Cy5.5, -APC, -V500 and -V450; CD4-APC, -BV421, -APC-Cy7 and -APC-H7; CD8a-APC-Cy7, -PE-Cy7 and -APC-H7; CD25-APC, -APC-Cy7 and -V450; CD44-PE; CD45-V500; CD45RB-PE; CD62 L-APC; CD69-APC; CD73-BV605 (clone TY/11.8) and PD1-PerCP-Cy5,5 (clone J43) were purchased by BD Biosciences (Germany). Purified anti-mouse CD3e and CD28, as well as FOXP3-FITC, CD73- eFluor® 450, -PE (clone TY/11.8 for both) and CD39-PE-Cy7, -PE (clone DM24S1 for both) were from eBioscience (Germany). BSA, Trypan blue, PMA, and Ionomycin were obtained from Sigma (Germany). Phosphate buffer saline (PBS), RPMI-1640 with L-glutamine and Penicillin-Streptomycin were purchased by PAA (Germany). Fetal bovine serum (FBS) was from PAN Biotech (Germany). Brefeldin A was purchased by BD Biosciences (Germany). Foxp3/Transcription Factor Staining Buffer Set and Permeabilization Buffer (10X) were obtained from eBioscience (Germany). Red Blood Cell (RBC) Lysis Buffer was from Biolegend (Germany).

Shevchenko I., Mathes A., Groth C., Karakhanova S., Müller V., Utikal J., Werner J., Bazhin A.V, & Umansky V. (2020). Enhanced expression of CD39 and CD73 on T cells in the regulation of anti-tumor immune responses. Oncoimmunology, 9(1), 1744946.

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Multiparameter Flow Cytometry Protocol

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Single cell suspensions from organs and tissues were stained with optimized antibody dilutions for 15 minutes at 4°C (Supplementary Table S4). Antibodies were directly conjugated to Brilliant Ultraviolet (BUV)395, Brilliant Violet (BV)786, BV711, BV650, BV605, Pacific Blue (PB), fluorescein isothiocyanate (FITC), phycoerythrin (PE), PE-Cy5.5, PE-Texas Red, peridinin-chlorophyll-protein complex (PerCP)-Cy5.5, PE-Cy7, allophycocyanin (APC), APC-Alexa 700, or biotin. Biotinylated antibodies were revealed with Streptavidin APCeFluor780. Cells were resuspended in FACS buffer (1% BSA+0.01% NaN₃ in PBS1x, filtered) and incubated with rat plus rabbit serum followed by incubation with antibody cocktail against surface molecules. For intracellular staining, cells were fixed (IC Fixation Buffer, eBioscience) and permeabilized (Permeabilization Buffer 10x, eBioscience). Dead cells were excluded through 4,6 diamidino-2-phenylindole (DAPI) uptake or Aqua live/dead staining. Doublets were excluded through forward scatter–height by forward scatter–width and side scatter–height by side scatter–width parameters. Cells were acquired on a LSR Fortessa SORP (BD Biosciences) and data were analyzed using FlowJo (Tree Star). Cell sorting was performed on a FACSAria II (BD Biosciences), and cell purity after sorting was > 99%.

Nath P.R., Pal-Nath D., Mandal A., Cam M.C., Schwartz A.L, & Roberts D.D. (2019). Natural killer cell recruitment and activation are regulated by CD47 expression in the tumor microenvironment. Cancer immunology research, 7(9), 1547-1561.

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