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Pe cy7 conjugated cd45

Manufactured by BioLegend
Sourced in United States

The PE-Cy7-conjugated CD45 is a flow cytometry reagent used to detect the CD45 antigen expressed on the surface of various hematopoietic cell types. It is a fluorescent-labeled monoclonal antibody that can be used to identify and analyze these cells in a sample.

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2 protocols using pe cy7 conjugated cd45

1

Quantifying C. albicans Infection in Mice

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Mice (WT and Gsdmd-/-) were intravenously challenged with 3 × 105 CFU WT (GFP+) or candidalysin-deficient (ece1Δ/Δ) C. albicans. Three days after C. albicans infection, kidneys were homogenized using 40 µm cell strainers. Red blood cells were lysed with 1 ml ACK lysis buffer (Gibco) for 5 min at room temperature. Cells were then stained with PE-Cy7-conjugated CD45 (BioLegend), PE-conjugated F4/80 (BioLegend), and APC-conjugated CD11b (BioLegend) antibodies for 30 min. For cells isolated from WT (GFP+) C. albicans-infected mice, macrophages containing trapped C. albicans were recognized by GFP fluorescence. For cells isolated from candidalysin-deficient (ece1Δ/Δ) C. albicans-infected mice, the samples were fixed and permeabilized (BD Biosciences) after surface marker staining, and then stained with FITC-conjugated anti-C. albicans antibody (thermofisher) for 1 h. Flow cytometry was performed on the LSRFortessa (BD Biosciences) instrument. Flow cytometry data were analyzed with FlowJo software.
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2

Isolation and Characterization of GFP-Labeled Mesenchymal Stem Cells from Synovium

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1 Â 10 6 GFP þ MSCs were injected 1week after the ACLT, then synovium was harvested 1day after the injection, following digestion with collagenase V for three hours. After filtering through a 70 mm cell strainer and centrifuging at 1500 rpm for 5 min, cells were suspended in ice-cold Hank's balances salt solution and then stained for 30 min on ice with a monoclonal antibody of APCconjugated CD90 (Becton, Dickinson and Company, Franklin Lakes, NJ, USA). Synovial fluid was also extracted from the same knee, and prepared in the same manner without collagenase digestion. Propidium iodide (PI) fluorescence was measured, and a live cell gate was defined that excluded the cells positive for PI. Additional gates were defined as positive for GFP and CD90. Double positive cells were further analyzed for PE-conjugated CD29, PEconjugated CD31, and PEcy7-conjugated CD45 (Biolegend, San Diego, CA, USA). Flow-cytometric analysis and sorting were performed on a MoFlo (Beckman Coulter, Brea, CA, USA), and the data were analyzed using FlowJo software (Tree Star, Ashland, OR, USA) 26 .
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