DPBS/BSA buffer: Prepare a solution containing Dulbecco’s phosphate-buffered saline (DPBS) with Ca2+ and Mg2+ and 0.5% bovine serum albumin (BSA) by diluting MACS® BSA Stock Solution 1:20 with DPBS. Keep buffer cold (2–8 °C). Prepare fresh on the day of use and degas the buffer, as air bubbles could block the column.
Coat a 24-well culture dish with 0.01% Poly-L-lysine overnight at 37 °C. Wash three times with ddH₂O the following day.
For cell culture, prepare the following medium: MACS Neuro Medium containing 2% MACS NeuroBrew®-21, 1% penicillin/streptomycin and 0.5 mM L-glutamine.
Dissociate mouse neonatal brain using the Neural Tissue Dissociation Kit – Postnatal Neurons. Follow the protocol of the kit data sheet.
Download data sheet
Staining buffer: Prepare a solution containing autoMACS® Running Buffer with FcR Blocking Reagent, mouse (1:10).
|Primary Antibody||Secondary Antibody|
|Antibody||Incubation temperature||Incubation time||Recommended antibody concentration||Secondary antibody||Incubation temperature||Incubation time|
|Anti-ACSA-2||Room temperature||10 minutes||1–5 µg/mL||Anti-rat IgG2b||Room temperature||10 minutes|
|Anti-GLAST (ACSA-1)||Room temperature||10 minutes||1–5 µg/mL||Anti-mouse IgG2a||Room temperature||10 minutes|
|Anti-O4||Room temperature||10 minutes||5–10 µg/mL||Anti-mouse IgM||Room temperature||10 minutes|
|Anti-PSA-NCAM||Room temperature||10 minutes||5–10 µg/mL||Anti-mouse IgM||Room temperature||10 minutes|
|CD11b||Room temperature||10 minutes||5–10 µg/mL||Anti-rat IgG2bκ||Room temperature||10 minutes|
|CD68||2–8 °C||Overnight||5–10 µg/mL||Anti-rat IgG2a||Room temperature||1 hour|
|CD171 (L1CAM)||2–8 °C||Overnight||5–10 µg/mL||Anti-rat IgG2a||Room temperature||1 hour|
|Column||Max. number of labeled cells||Max. number of total cells||Selector|
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