Red blood lysing solution

Manufactured by Miltenyi Biotec

Red Blood Lysing Solution is a buffer solution designed for the lysis of red blood cells. It effectively removes red blood cells from a cell suspension, allowing for the isolation and analysis of other cell types.

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2 protocols using red blood lysing solution

Immunophenotyping of Whole Blood in GCA

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Venous blood was drawn from GCA patients into heparin-containing tubes. Whole blood immunophenotyping was performed using 7-Color Immunophenotyping kit with the following antibodies (Miltenyi Biotec, catalog #130-098-456): CD14-FITC (clone Tük4), CD56-PE (clone REA196), CD16-PE (clone REA423), CD4-PerCP (clone VIT4), CD19-PE-Vio® 770 (clone LT19), CD3-APC (clone BW264/56), CD8-APC-Vio 770 (clone BW135/80), CD45-VioBlue® (clone 5B1). Briefly, 100 μl of whole blood was incubated with 10 μl immunophenotyping reagent for 10 min in the dark, at 4°C. After incubation, whole blood was lysed using Red Blood Lysing Solution (Miltenyi Biotec, catalog #130-098-456). Neutrophil phenotyping was performed in 50 μl of whole blood, incubated for 30 min at 4°C in the dark, with the following antibodies (eBioscience): CD16-PE (clone eBioCB16; catalog #50-112-4738), CD62L-PE-Cy5 (clone DREG56; catalog #50-140-71) and CD11b-APC (clone ICRF44; catalog #17-0118-42). After incubation, samples were lysed, using Whole Blood Lysing Reagent Kit (Beckman Coulter; catalog #6602764). All samples were analyzed using flow cytometer MACSQuant Analyzer 10 (Miltenyi Biotec). Analysis of flow cytometry data was performed using MACSQuantify (Analysis Software version 2.8, Miltenyi Biotec) and FlowLogic (Flow Cytometry Analysis Package, version 7.00.0a, Invasion Software Technologies Pvt Ltd).

Kuret T., Frank-Bertoncelj M., Lakota K., Žigon P., Thallinger G.G., Kopitar A.N., Čučnik S., Tomšič M., Hočevar A, & Sodin-Šemrl S. (2022). From Active to Non-active Giant Cell Arteritis: Longitudinal Monitoring of Patients on Glucocorticoid Therapy in Combination With Leflunomide. Frontiers in Medicine, 8, 827095.

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Immunophenotyping of Neutrophils in IgAV Patients

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Venous blood was drawn from IgAV patients and HBDs and collected into heparin-containing tubes. Whole blood immunophenotyping was performed using 7-Color Immunphenotyping kit (Miltenyi Biotec). Briefly, 100 μl of whole blood was incubated with 10 μl immunophenotyping reagent for 10 min in the dark, at 4 °C. After incubation, erythrocytes were lysed using Red Blood Lysing Solution (Miltenyi Biotec). Neutrophil phenotyping was performed in 50 μl of whole blood, incubated for 30 min at 4 °C in the dark with mouse anti-human antibodies to CD16 (conjugated to PE, clone eBioCB16, eBioscience), CD62L (conjugated to PE-Cyanine 5, clone DREG56, eBioscience), and CD11b (conjugated to APC, clone ICRF44, eBioscience). After incubation, samples were lysed, using Whole Blood Lysing Reagent Kit (Beckman Coulter). All samples were analyzed using flow cytometer MACSQuant Analyzer 10 (Miltenyi Biotec). Analysis of flow cytometry data was performed using MACSQquantify (Analysis Software version 2.8) and FlowLogic (Flow Cytometry Analysis Package, version 700.0a).

Kuret T., Lakota K., Žigon P., Ogrič M., Sodin-Šemrl S., Čučnik S., Tomšič M, & Hočevar A. (2019). Insight into inflammatory cell and cytokine profiles in adult IgA vasculitis. Clinical rheumatology, 38(2).

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