Fix and perm a and b

Manufactured by Thermo Fisher Scientific

Fix and Perm A and B are laboratory fixation and permeabilization reagents used in cell and tissue sample preparation for various analytical techniques. They are designed to preserve the structural integrity of samples while allowing access to intracellular components for subsequent analysis. The core function of these products is to enable effective fixation and permeabilization of samples without making claims about their intended use.

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Lab products found in correlation

Anti ifn γ apc, by BD (2 mentions) Anti cd107a phycoerythrin pe cy5, by BD (2 mentions) Anti cd3 alexafluor700, by BD (2 mentions) Anti mip1β pe, by BD (2 mentions) Anti cd16 allophycocyanin apc cy7, by BD (2 mentions) Brefeldin a, by Merck Group (2 mentions) Golgistop, by BD (2 mentions) Rosettesep, by STEMCELL (2 mentions) Anti cd56 pe cy7, by BD (2 mentions)

Close spelling variants of this product

FIX & PERM medium A and B Fix and Perm media A and B Fix and Perm A and B solutions Perm A and B Fix and Perm Fix/Perm Perm B

2 protocols using fix and perm a and b

NK Cell Activation Assay Protocol

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An assay to determine NK cell activation (NKA) state based on the expression of surface CD107a and intracellular production of IFNγ and MIP1β was performed as previously described. NK cells were isolated from whole blood from healthy donors using negative selection with RosetteSep (STEMCELL Technologies). Following a pulse with SF162 gp120 (60 μg/ml), T lymphoblast CEM‐NKr cells and isolated primary NK cells were mixed at a ratio of 1:5, and purified IgG, anti‐CD107a‐phycoerythrin (PE)‐Cy5 (BD), brefeldin A (10 mg/ml) (Sigma), and GolgiStop (BD) were added. After a 5‐h incubation at 37°C, cells were first stained for surface markers using anti‐CD16‐allophycocyanin (APC)‐Cy7 (BD), anti‐CD56‐PE‐Cy7 (BD), and anti‐CD3‐Alexa Fluor 700 (BD) and then stained intracellularly with anti‐IFNγ‐APC (BD) and anti‐MIP1β‐PE (BD) after treatment with Fix and Perm A and B solutions (Invitrogen). Cells were then fixed in 4% paraformaldehyde and analyzed by flow cytometry. NK cells were defined as CD3‐negative and CD16‐positive and/or CD56‐positive, and the percent of NK cells positive for each marker was determined.

Alter G., Dowell K.G., Brown E.P., Suscovich T.J., Mikhailova A., Mahan A.E., Walker B.D., Nimmerjahn F., Bailey‐Kellogg C, & Ackerman M.E. (2018). High‐resolution definition of humoral immune response correlates of effective immunity against HIV. Molecular Systems Biology, 14(3), e7881.

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NK Cell-Mediated Immune Function Assay

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An assay to determine the expression of surface CD 107a and intracellular production of IFN-γ and MIP-1β was performed by pulsing the CEM-NKr CCR5⁺ T lymphoblast cell line with rgp120SIV_mac251 (60 mg/ml), as previously described^{51 (link)}. NK cells were isolated from whole blood from healthy donors using negative selection with RosetteSep (STEMCELL Technologies), as recommended by the manufacturer. The CEM-NKr cells and isolated primary NK cells were mixed at a ratio of 1:5, and purified IgG, anti- CD107a-phycoerythrin (PE)-Cy5 (BD), brefeldin A (10 mg/ml) (Sigma-Aldrich), and GolgiStop (BD) were added for 5 h at 37 °C. The cells were then first stained for surface markers using anti-CD 16-allophycocyanin (APC)-Cy7 (BD), anti-CD56-PE-Cy7 (BD) and anti-CD3-Alexa Fluor 700 (BD) and then stained intracellularly with anti-IFN-γ-APC (BD) and anti-MIP-1β-PE (BD) using Fix and Perm A and B solutions (Invitrogen). The cells were then fixed in 4% paraformaldehyde and analyzed using flow cytometry. NK cells were defined as CD3⁻ and CD16⁻ and/or CD56⁺.

Vaccari M., Gordon S.N., Fourati S., Schifanella L., Liyanage N.P., Cameron M., Keele B.F., Shen X., Tomaras G.D., Billings E., Rao M., Chung A.W., Dowell K.G., Bailey-Kellogg C., Brown E.P., Ackerman M.E., Vargas-Inchaustegui D.A., Whitney S., Doster M.N., Binello N., Pegu P., Montefiori D.C., Foulds K., Quinn D.S., Donaldson M., Liang F., Loré K., Roederer M., Koup R.A., McDermott A., Ma Z.M., Miller C.J., Phan T.B., Forthal D.N., Blackburn M., Caccuri F., Bissa M., Ferrari G., Kalyanaraman V., Ferrari M.G., Thompson D., Robert-Guroff M., Ratto-Kim S., Kim J.H., Michael N.L., Phogat S., Barnett S.W., Tartaglia J., Venzon D., Stablein D.M., Alter G., Sekaly R.P, & Franchini G. (2016). Adjuvant-dependent innate and adaptive immune signatures of risk of SIVmac251 acquisition. Nature medicine, 22(7), 762-770.

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