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Perm wash buffer

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Perm/Wash buffer is a laboratory reagent used to prepare cells for analysis. It is designed to permeabilize cell membranes and wash the cells prior to further processing or staining.

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Lab products found in correlation

Fc blocker, by BD (2 mentions) Fix perm buffer, by R&D Systems (2 mentions) Pd l1, by BD (2 mentions) Cytoflex, by Beckman Coulter (2 mentions) Annexin 5, by BD (2 mentions) F4 80, by BD (2 mentions) Perm wash buffer, by BD (2 mentions) Perm wash buffer, by Beckman Coulter (1 mentions) Lightning link antibody labeling kit, by Novus Biologicals (1 mentions) Lsrii cytometer, by BD (1 mentions) Lsr 2 flow cytometer, by BD (1 mentions)

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Wash buffer (16 citations)

4 protocols using perm wash buffer

1

Multicolor Flow Cytometry for Immune Profiling

Cited 2 times
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For cell surface staining, cells were washed with phosphate-buffed saline (PBS) and blocked with Fc blocker (BD Biosciences, San Jose, California, USA). Fluorochrome-labeled antibodies (Annexin-V, CD45, CD11c, CD4, CD8, CD11b, Ly6G, Ly6C, PD-1, PD-L1, F4/80, CD56, CD86, HLA-DR, CD206, and CD44) were obtained from BD Biosciences (Franklin Lakes, USA), added, and stained for 30 min as described.54 (link) For intracellular staining, cells were permeabilized with Fix/Perm buffer (#FC009, R&D Systems, Minneapolis, Minnesota, USA) for 20 min and then washed with Perm/Wash buffer (R&D Systems). Fluorochrome-labeled antibodies (IFNγ and TNFα) (#562019, #561062, BD Biosciences) were diluted in Perm/Wash buffer and stained for 30 min as described.55 (link) All samples were analyzed on a CytoFlex flow cytometer (Beckman Coulter, California, USA).
Hong B., Sahu U., Mullarkey M.P., Hong E., Pei G., Yan Y., Otani Y., Banasavadi-Siddegowda Y., Fan H., Zhao Z., Yu J., Caligiuri M.A, & Kaur B. (2023). PKR induces TGF-β and limits oncolytic immune therapy. Journal for Immunotherapy of Cancer, 11(2), e006164.
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2

Multiparametric Islet Cell Analysis

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EP12-BTMR-X stained intact islets were dispersed with 0.25% trypsin for 10 min at 37 °C and fixed with 4% paraformaldehyde for 20 min and subsequently permeabilized with perm/wash buffer (BD). Intracellular staining was carried out in perm/wash buffer containing 2% rat serum with anti-insulin APC (R&D systems) and anti-glucagon (R&D Systems) conjugated to PerCP-Cy5.5 using the Lightning-link antibody labeling kit (Novus Biologicals) for 2 h at RT. Following extensive washing, cells were filtered and acquired on a LSRII cytometer (BD). Data was analyzed using FlowJo software (Tree Star).
Clardy S.M., Mohan J.F., Vinegoni C., Keliher E.J., Iwamoto Y., Benoist C., Mathis D, & Weissleder R. (2015). Rapid, high efficiency isolation of pancreatic ß-cells. Scientific Reports, 5, 13681.
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3

Comprehensive Cell Surface and Intracellular Staining

Cited 1 time
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For cell surface staining, cells were washed with PBS and blocked with Fc blocker (BD Biosciences, San Jose, CA). Fluorochrome labeled antibodies (Annexin-V, CD45, CD11c, CD4, CD8, CD11b, Ly6G, Ly6C, PD-1, PD-L1, F4/80, CD56, CD86, HLA-DR, CD206, CD44) were obtained from BD Biosciences (Franklin Lakes, USA), added and stained for 30 min as described (9 (link)). For intracellular staining, cells were permeabilized with Fix/Perm buffer (FC009, R&D systems) for 20min and then washed with Perm/Wash buffer (R&D systems, Minneapolis, MN). Fluorochrome labeled antibody Foxp3 (560408, BD Biosciences) were diluted in Perm/Wash buffer and stained for 30min as described (10 (link)). All samples were analyzed on a CytoFlex flow cytometer (Beckman Coulter, CA).
Hong B., Chapa V., Saini U., Modgil P., Cohn D.E., He G., Siddik Z.H., Sood A.K., Yan Y., Karuppaiyah S., Pei G., Zhao Z., Yoo J.Y, & Kaur B. (2020). Oncolytic HSV therapy modulates vesicular trafficking inducing cisplatin sensitivity and anti-tumor immunity. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(2), 542-553.
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4

Flow Cytometric Analysis of Insulin Expression

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For flow cytometry, cells were seeded into 24 well plates at a density of 0.5 million/cm2. On the second day, for each well, cells were trypsinated and washed once with PBS. Cells were fixed in 1% formaldehyde (Sigma) for 20 minutes and permeabilized in Perm/ Wash buffer (BD Biosciences) for 40 minutes at room temperature. Cells were then stained with Insulin antibodies, rat IgG (R&D Systems) at 1:100 dilution in 50 ul Perm/Wash buffer in 4°C overnight. On the second day, cells were washed once with Perm/Wash buffer and stained with PE-conjugated anti-rat IgG secondary antibodies at 1:200 dilution in 50 ul Perm/Wash buffer at room temperature for 30 minutes in the dark. Cells were then washed once with PBS and analyzed in BD LSR II Flow Cytometer (BD Biosciences).
Fang Z., Weng C., Li H., Tao R., Mai W., Liu X., Lu L., Lai S., Duan Q., Alvarez C., Arvan P., Wynshaw-Boris A., Li Y., Pei Y., Jin F, & Li Y. (2019). Single-Cell Heterogeneity Analysis and CRISPR Screen Identify Key β-Cell-Specific Disease Genes. Cell reports, 26(11), 3132-3144.e7.
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