The three Sham group specimens, the three CLP group specimens, and the three Maresin1 group specimens were mixed, respectively, for single-cell RNA sequencing. Single-cell suspensions of lung tissue specimens were prepared according to OE Biotech Co., Ltd.’s advice [22 (link)]. The cells were then sorted using Miltenyi mouse CD45 magnetic beads according to the manufacturer’s recommendations and sequenced at a 1:1 ratio. According to the manufacturer’s guidelines, the single-cell libraries were constructed by 10× Genomics Chromium Single Cell 3ʹ Reagent Kits v3.1. The Illumina Nova 6000 PE150 platform was used to sequence the libraries.
Mouse cd45 magnetic beads
Manufactured by Miltenyi Biotec
Mouse CD45 magnetic beads are a laboratory reagent used for the isolation and enrichment of mouse CD45-positive cells from a heterogenous cell population. The beads are coated with antibodies that bind specifically to the CD45 surface antigen expressed on mouse leukocytes.
Automatically generated - may contain errors
3 protocols using mouse cd45 magnetic beads
1
Single-cell RNA sequencing of lung tissue
2
Mouse Cell Depletion from BM
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Total BM from NSGS mice was depleted of mouse cells with mouse CD45 magnetic beads (Miltenyi Biotec; catalog no. 130-052-301, RRID:AB_2877061) and negatively selected human cells were used.
3
Leukemic Blast Cell Cycle Analysis
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Cell labeling was done by i.p. injection of mice with 50 mg/kg of freshly prepared EdU (Cayman Chemical Company, catalog no. 20518). After three to four hours, BM was harvested, and human cells were negatively selected by mouse cell depletion using mouse CD45 magnetic beads (Miltenyi). The human BM cells were then stained with CD45 and CD33 to identify the leukemic blast. Cell-cycle/proliferation analysis was performed using the Click-iT Plus EdU Flow Cytometry Assay Kit (Invitrogen, catalog no. C10420) following the manufacturers’ instructions. Fixable viability dye (BioLegend) was used to discriminate the death population. Single-color controls were used to set compensations, and fluorescence minus one control were used to set gates. Analysis was performed with FlowJo software.
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