This protocol has been developed to generate highly purified and viable endothelial cells from adult mouse brain tissue. Brain tissue from mice older than P7 is dissociated into single-cell suspensions using the Adult Brain Dissociation Kit and the gentleMACS™ Octo Dissociator with Heaters for mechanical dissociation during on-instrument enzyme incubation. After dissociation, the myelin and cell debris is removed using the Debris Removal Solution and is followed by subsequent removal of erythrocytes using the Red Blood Cell Removal Solution. Endothelial cells are enriched by depletion of CD45+ cells with CD45 MicroBeads followed by a positive selection using CD31 MicroBeads.
Preparation of enzyme mix
|Enzyme mix 1||Enzyme mix 2|
Preparation of 1x Red Blood Cell Removal Solution
Coat the cell culture dishes with Human Fibronectin (Fragment). The coating concentration should be 3 μg of Human Fibronectin per cm2.
For example, when coating a 96-well plate (0.32 cm2 per well):
|Debris Removal Solution||D-PBS||Overlay (D-PBS)|
|900 µL||3100 µL||4 mL|
|1800 µL||6200 µL||4 mL|
Magnetic separation with LD Columns
Magnetic separation with the autoMACS® Pro Separator
Magnetic separation with MS Columns
Magnetic separation with the autoMACS Pro Separator
Unless otherwise specifically indicated, Miltenyi Biotec products and services are for research use only and not for therapeutic or diagnostic use.
If not otherwise specified, all marks displayed on the Miltenyi Biotec Internet site and documents are subject to the trademark rights of Miltenyi Biotec, including each of Miltenyi Biotec's primary brands, its model name plates, and its corporate logos and emblems.
For copyright note and a complete list of trademarks visit www.miltenyibiotec.com/legalnotes.
|Column||Max. number of labeled cells||Max. number of total cells||Separator|
|MS||1x10⁷||2x10⁷||MiniMACS™, OctoMACS™, |
|LD||2x10⁷||4x10⁷||MidiMACS™, OctoMACS™, |
▲ Note: Column adapters are required to insert certain columns into the SuperMACS II Separator. For details refer to the respective MACS Separator data sheet.
Seems like you are coming from USA!
Do you want to visit our website in your country?