After digestion of the hearts using the method for CM isolation, noncardiomyocyte cells were filtered through 100- and 40-mm cell strainers (VWR, Radnor, PA) and centrifuged at 134 × g for 10 min. Cells were resuspended in 400 μl of PBS and 100 μl was taken and incubated with the following Abs at the various dilutions listed: 1:2000 CD45 PE (catalog no. 553081; BD Biosciences), 1:100 Ly6G BV421 (catalog no. 562737; BD Biosciences), 1:1000 Ly6C PECy7 (catalog no. 25-5932-80; eBioscience), 1:100 F4/80 Alexa Fluor 647 (catalog no. 565853; BD Biosciences), and 1:1000 Zombie NIR (catalog no. 423105; BioLegend). The samples were then fixed with 1% paraformaldehyde and run on a BD LSR II.
100 and 40 mm cell strainers
Manufactured by Avantor
The 100- and 40-mm cell strainers are laboratory equipment designed for filtering and separating cells from a suspension. They feature a mesh screen that allows the passage of single cells while retaining larger cell aggregates, debris, and tissue fragments.
Automatically generated - may contain errors
2 protocols using 100 and 40 mm cell strainers
1
Isolation and Characterization of Cardiac Immune Cells
2
Isolation and Characterization of Cardiac Immune Cells
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