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Anti cd45 magnetic beads conjugated antibody

Manufactured by Miltenyi Biotec

Anti-CD45 magnetic beads-conjugated antibody is a laboratory reagent used for the isolation and enrichment of CD45-positive cells from complex biological samples. The antibody is coupled to magnetic beads, allowing for efficient separation of the target cells using a magnetic field.

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2 protocols using anti cd45 magnetic beads conjugated antibody

1

CD45+ Cell Isolation from Samples

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SVCs were resuspended in 500 mL of magnetic bead buffer (MBB) consisting of PBS without calcium and magnesium, 0.5% w/v bovine serum albumin (BSA), and 2 mM ethylenediaminetetraacetic acid (EDTA). The cell suspension was then filtered through a 30-mm pre-separation filter (Miltenyi, Bergisch Gladbach, Germany) following three filter washes to remove any large particles and debris.
The resulting cell suspension was then separated at 300 × g for 5 min at 4°C and resuspended in 90 mL of MBB along with 10 mL of anti-CD45 magnetic beads-conjugated antibody (Miltenyi) for each 10ˆ7 cell sample. The cell suspension was incubated for 15 min at 4°C, then diluted with 2 mL of MBB and centrifuged. The resulting cell pellet was resuspended in 500 mL of MBB, applied onto hydrated MS-columns (Miltenyi), washed three times with 500 mL of MBB, and collected with 1 mL of MBB through piston elution.
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2

Isolation of Stromal Vascular Cells and Mitochondria

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Stromal vascular cells were resuspended in 500 mL of magnetic bead buffer (MBB) (PBS 1× without calcium and magnesium, 0.5% w/v BSA, and 2 μM ethylenediaminetetraacetic acid [EDTA]) and filtered through a 30-μM pre-separation filter (Miltenyi, Bergisch Gladbach, Germany) following three filter washes to remove large particles and debris. The cell suspension was separated at 300 × g for 5 min at 4°C and resuspended in 90 mL of MBB plus 10 mL of anti-CD45 magnetic beads-conjugated antibody (Miltenyi) for each 107 cell sample. After 15 min of incubation at 4°C, the cell suspension was diluted with 2 mL of MBB and centrifuged. The cell pellet was resuspended in 500 mL of MBB, applied on hydrated MS-columns (Miltenyi), washed three times with 500 mL of MBB, and collected with 1 mL of MBB through piston elution. Incubation with anti-CD11b and anti-F4/80 magnetic beads-conjugated antibodies (Miltenyi) was performed in a unique incubation, as was performed for CD45. BAT was homogenized using a Potter–Elvehjem polytetrafluoroethylene pestle and a glass tube and TO μM 22+ mitochondria were magnetically isolated by a commercially available kit (Miltenyi).
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