The application protocol was developed to isolate high yields of viable endothelial cells from mouse neonatal brain tissue. Cells can be cultured or analyzed by flow cytometry afterwards.
PBE buffer: Prepare a solution containing phosphate-buffered saline (PBS), pH 7.2, 0.5% bovine serum albumin (BSA), and 2 mM EDTA by diluting MACS BSA Stock Solution 1:20 with autoMACS® Rinsing Solution. Keep buffer cold (2−8 °C).
Coat the necessary 96-well culture dishes with 100 μg/mL fibronectin overnight and in an incubator at 37 °C.
Download data sheet
For a detailed immunofluorescent staining protocol, refer to the data sheet of the chosen CD31 antibodies.
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▲ Note: Make sure the antigen epitope that is necessary for downstream applications is conserved during the dissociation procedure.
For a detailed list of antigen compatibilities and the right choice of NTDK refer to the table on NTDK product page at www.miltenyibiotec.com.
In case your epitope of interest is not listed please contact technical support. You can also perform a staining experiment with this antibody after using different enzyme concentrations, i.e., different dilutions of Enzyme P or T (e.g., for NTDK (P) 1:5, 1:10; for NTDK (T) 1:2.5) prior to isolation experiments to analyze the stability of your antibody epitope.