For RNA-seq, qPCR, immunoblotting, and immunoprecipitation experiments neural progenitor cells (NPCs) from 3.5DDC and onward were isolated from differentiating ESCs cultures by Prominin1-based magnetic sorting (p-MACS) according to the manufacturer’s instructions (Miltenyi Biotec). Briefly, cells were dissociated using TrypLE Express, washed twice with ice-cold PBS supplemented with 0.5% bovine serum albumin and 2 mM EDTA (Sigma-Aldrich), and passed through a 40-μm cell strainer. Dissociated cells were incubated with magnetic beads conjugated with anti–prominin-1 antibody. Prominin1+ cells were separated on magnetic columns and processed for biochemical analysis. To assess purity of sorted fraction, cells were plated, incubated for 2 hours, and fixed for immunocytochemistry.
P macs
Manufactured by Miltenyi Biotec
The P-MACS is a lab equipment product offered by Miltenyi Biotec. It is a magnetic cell separation system designed for the isolation and purification of specific cell populations from complex biological samples.
Automatically generated - may contain errors
2 protocols using p macs
1
Prominin1-based Magnetic Sorting of NPCs
2
Differentiation of mESCs into Neural Progenitors
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E14 mESCs were differentiated as described before (43 (link)).Total RNA was isolated from cells every 24 h between 0 and 3 days of differentiation condition (DDC) using the ZymoPure™ kit according to manufacturer's instructions. Three DDC cultures were enriched for neural progenitors using prominin1-based magnetic activated cell sorting (p-MACS) (Miltenyi Biotec).
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