Miltenyi de myelination kit

Manufactured by Miltenyi Biotec

The Miltenyi de-myelination kit is a laboratory product designed to assist in the removal of myelin from biological samples. Its core function is to facilitate the isolation and separation of cells from myelinated tissue, enabling further analysis and research.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions) Rnase inhibitor, by Takara Bio (2 mentions) Countess automated cell counter, by Thermo Fisher Scientific (2 mentions) Adult brain dissociation kit, by Miltenyi Biotec (2 mentions) Facsaria flow cytometer, by BD (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by BD (2 mentions) Clone hi30, by BD (1 mentions)

2 protocols using miltenyi de myelination kit

Isolation of Brain Immune Cells

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Fresh autopsy tissue specimens from DLPFC (Brodmann Area 10) were placed in tissue preservation buffer (Miltenyi Biotech) and stored at 4°C for ≤ 48hrs before processing using the Adult Brain Dissociation Kit (Miltenyi Biotech cat.# 130107677), according to the manufacturer’s instructions. RNase inhibitors (Takara Bio) were used throughout the cell prep. Following de-myelination (Miltenyi de-myelination kit, Miltenyi Biotech, cat.# 130096733) cells were incubated in antibody (CD45: BD Pharmingen, Clone HI30 and CD11b: BD Pharmingen, Clone ICRF44) at 1:500 for 1 hour in the dark at 4°C with end-over-end rotation. Prior to fluorescence activated cell sorting (FACS), DAPI (Thermoscientific) was added at 1:1000 to facilitate selection of viable cells. Viable (DAPI⁻) CD45⁺CD11b⁺ cells were isolated by FACS using a FACSAria flow cytometer (BD Biosciences). Following FACS, cellular concentration and viability was confirmed using a Countess automated cell counter (Life technologies).

Roussos P., Kosoy R., Fullard J., Bendl J., Kleopoulos S., Shao Z., Argyriou S., Mathur D., Vicari J., Ma Y., Humphrey J., Brophy E., Raj T., Katsel P., Voloudakis G., Lee D., Bennett D., Haroutunian V, & Hoffman G. (2024). Alzheimer’s disease transcriptional landscape in ex-vivo human microglia. Research Square.

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Isolation of Brain-Derived Immune Cells

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Fresh autopsy and biopsy tissue specimens were processed using the Adult Brain Dissociation Kit (Miltenyi Biotech cat.# 130107677), according to manufacturer’s instructions. Following de-myelination (Miltenyi de-myelination kit, Miltenyi Biotech, cat.# 130096733) cells were incubated in antibody (CD45: BD Pharmingen, Clone HI30 and CD11b: BD Pharmingen, Clone ICRF44) at 1:500 for 1 hour in the dark at 4 °C with end-over-end rotation. RNase inhibitors (Takara Bio) were used throughout cell preparation. Prior to fluorescence-activated cell sorting (FACS), DAPI (Thermoscientific) was added at 1:1,000 to facilitate the separation of live from dead cells. Viable (DAPI⁻) CD45⁺ cells were isolated via FACSAria flow cytometer (BD Biosciences). Following FACS, cell concentrations and viability were confirmed using a Countess automated cell counter (Life technologies).

Kosoy R., Fullard J.F., Zeng B., Bendl J., Dong P., Rahman S., Kleopoulos S.P., Shao Z., Girdhar K., Humphrey J., de Paiva Lopes K., Charney A.W., Kopell B.H., Raj T., Bennett D., Kellner C.P., Haroutunian V., Hoffman G.E, & Roussos P. (2022). Genetics of the human microglia regulome refines Alzheimer’s disease risk loci. Nature genetics, 54(8), 1145-1154.

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