Resource Summary Report  New Search Previous Search Results

URL: https://dx.doi.org/10.17504/protocols.io.7xjhpkn

Authors: Sam Li

Group: BioLegend

Summary: Product description and procedure summary: The cells targeted by the Nanobeads are either selected or depleted by incubating your sample with the directly conjugated magnetic particles. The magnetically labeled fraction is retained by the use of a magnetic separator. After collection of the targeted cells, downstream applications include functional assays, gene expression, phenotypic characterization, etc. Note: This procedure is optimized for the isolation of 107 to 2 x 108 cells per tube. If working with fewer than 107 cells, keep volumes as indicated for 107 cells. For best results, optimize the conditions to your specific cell number and tissue. Prepare fresh MojoSort™ Buffer solution by diluting the 5X concentrate with sterile distilled water.Scale up volumes if using 14mL tubes and Magnet, and place the tube in the magnet for 10 minutes. Sample Preparation: Enzymatic digestion of mouse spleen is recommended to achieve the highest purity and yield of CD11c+ cells. There are several protocols published that can be applied. As a general guideline, cut mouse spleen into pieces and incubate in 0.5 mg/ml Collagenase for 30 to 60 minutes at room temperature or 37°C. Place the tube in a rocking platform with continuous agitation or gently pipette every 10 minutes. Alternatively, inject 1 ml of enzymes solution in the uncut organ. Force the tissue through a 70µm filter to prepare a single cell suspension, and wash with complete media. Resuspend cells in 0.1 mg/ml DNase 1 solution and incubate at room temperature for 10 minutes. Again, filter cells through a 70 µm filter and wash with complete media. Resuspend in complete media or MojoSort™ Buffer and keep on ice until ready to use.

Affiliations: BioLegend

External URL: https://www.biolegend.com/protocols/mojosort-mouse-cd11c-nanobeads-protocol/4401/

Version: 1

Publication Date: 2019

Expand All Collapse