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Macsquant

Manufactured by FlowJo
Sourced in Netherlands

The MACSQuant is a compact and versatile flow cytometer designed for cell analysis and sorting. It offers precise measurements of multiple parameters for a wide range of cell types, including fluorescently labeled cells. The MACSQuant provides reliable and reproducible results, making it a useful tool for various applications in life science research and clinical settings.

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2 protocols using macsquant

1

Evaluating Leukemia Cell Resistance to Momelotinib and Ruxolitinib

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Cell-intrinsic resistance towards momelotinib and ruxolitinib (Selleck Chemicals, Kirby Drive, Houston, USA) was evaluated as described previously [52 (link)]. Briefly, leukemic cells were exposed to a concentration range (24μM to 750 nM) of these compounds for four days and cytotoxicity was quantified using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). For TSLP stimulation, cells were pre-incubated for 1 hour with 25 ng/ml TSLP (R&D systems, Oxon, UK). In addition to single cell cultures, leukemic cells (1*106 cells) were co-cultured with primary MSCs (5*104 cells) for four days in a 24 well plate in the presence of a dilution series of momelotinib and ruxolitinib. Cell survival was quantified using flow cytometry (MACSQuant, FlowJo 10.0.8r1), and cells were stained with Brilliant Violet 421 anti-human CD19 antibody (Biolegend), FITC Annexin V (Biolegend), and Propidium Iodide (PI; Invitrogen, Bleiswijk, Netherlands), as described previously [45 (link)].
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2

Isolation of Skin Cells from Ear Samples

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Ear skin samples were digested in 1 mL of Dulbecco modified Eagle medium containing 20 mmol/L HEPES, 0.025 mg/mL Liberase, 396 U/mL DNase I, and 0.5 mg/mL hyaluronidase at 378C at 1400 rpm for 1 hour to isolate skin cells. Samples were passed through a 40-mm sieve, and the cell suspension was washed twice with PBS. For flow cytometry, skin or LN cells were resuspended in PBS/2% BSA. Cells (2-5 3 10 5 ) were stained with a mAb in 100 mL of staining solution for 30 minutes at 48C. Cell suspensions were washed twice and resuspended in 200 mL of PBS/2% BSA. For fluorescence-activated cell sorting analysis of intracellular cytokines, cells were incubated with Brefeldin A in RPMI 1640 with 10% FCS and 1% penicillin/streptomycin before fixation in forkhead box protein 3 Staining Buffer (eBioscience, San Diego, Calif) and staining. Analysis was done with the Miltenyi MacsQuant flow cytometer with MacsQuant or FlowJo analysis software (FlowJo, Ashland, Ore).
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