Fix and perm medium b

Manufactured by BD

Fix and Perm Medium B is a laboratory reagent used for tissue fixation and permeabilization. It serves to preserve cellular structures and facilitate access to intracellular components during sample preparation for analysis.

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Lab products found in correlation

Facsaria fusion flow cytometer, by BD (2 mentions) Fix and perm medium a, by Thermo Fisher Scientific (2 mentions) Fix and perm medium b, by Thermo Fisher Scientific (2 mentions) Ionomycin, by Merck Group (2 mentions) Phorbol myristate acetate (pma), by Merck Group (2 mentions) Golgiplug, by BD (2 mentions)

2 protocols using fix and perm medium b

Multiparametric Analysis of CNS-Derived Cells

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Cells isolated from the CNS were stimulated with PMA (500 ng/mL; Sigma Aldrich, St. Louis, MO), ionomycin (50 ng/mL; Sigma Aldrich, St. Louis, MO), and 1 μL/mL Golgiplug (BD Biosciences, Franklin Lakes, NJ) for 4 h at 37°C. Cells were washed with PBS containing 3% FBS. Surface antigens were stained with Abs in 100 μL of PBS/3% FBS for 20 min at 4°C. Cells were then fixed with 100 μL Fix and Perm Medium A (Thermo Fisher, Waltham, MA) for 20 min at room temperature and then permeabilized with Fix and Perm Medium B (Thermo Fisher, Waltham, MA) and stained with Abs against intracellular antigens in 100 μL Fix and Perm Medium B and 100 μL PBS/3%FBS for 1 h. Cells were then washed twice, resuspended in 500 μL PBS and analyzed on a BD FACSAria Fusion flow cytometer (BD Biosciences, Franklin Lakes, NJ).

Hwang D., Boehm A., Rostami A., Zhang G.X, & Ciric B. (2021). Oral D-mannose treatment suppresses experimental autoimmune encephalomyelitis via induction of regulatory T cells. Journal of neuroimmunology, 362, 577778.

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Flow Cytometric Analysis of Stimulated Cells

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Isolated cells were stimulated with PMA (500 ng/mL; Sigma Aldrich), ionomycin (50 ng/mL; Sigma Aldrich), and 1 µL/mL Golgiplug (BD Biosciences) for 4 h at 37 o C. After stimulation, cells were washed with PBS containing 3% FBS (v/v). Cell surface antigens were stained with Abs in 100 µL of PBS/3% FBS for 20-30 min at 4 o C. Cells were then washed and fixed with 100 µL Fix and Perm Medium A (Thermo Fisher) for 20 min at room temperature and washed again. Cells were permeabilized with Fix and Perm Medium B (Thermo Fisher) and stained with Abs against intracellular antigens in 100 µL Fix and Perm Medium B and 100 µL PBS/3%FBS for 1 h. Cells were then washed twice, resuspended in 500 µL PBS and analyzed on a BD FACSAria Fusion flow cytometer (BD Biosciences).

Hwang D., Seyedsadr M., Ishikawa L.L.W., Boehm A., Sahin Z., Casella G., Jang S., Gonzalez M.V., Garifallou J., Hákonarson H., Zhang W., Xiao D., Rostami A., Zhang G., & Ćirić B. (2020). CSF-1 maintains pathogenic but not homeostatic myeloid cells in the central nervous system during autoimmune neuroinflammation.

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