Immunoaffinity isolation of uEVs was performed using a commercially available kit (Exosome Isolation Kit Pan, human, Miltenyi Biotec, Bergisch Gladbach, Germany), according to the manufacturer’s instructions. The kit is based on the immunomagnetic isolation of uEVs carrying any of the surface epitopes CD9, CD63, or CD81. Briefly, pre-processed void urine was pre-cleared by centrifugation at 10.000× g for 30 min (4 °C). Then, 10 mL of the supernatant were concentrated using an Amicon Ultra-15 100 kDa filter (regenerated cellulose; Merck/Millipore) by centrifuging at 4.000× g for 10 min. The concentrate was adjusted to 2 mL with 0.9% NaCl. Then, 50 µL of antibody-loaded magnetic beads were added. After incubation for 1 h at room temperature, the sample was loaded onto an equilibrated separation column within a magnetic stand (MACS MultiStand with µMACS separator, Miltenyi Biotec). After serial washing steps, the column was removed from the magnetic stand. Then, uEV-bead aggregates were eluted by flushing with 100 µL of the supplied isolation buffer. The samples were adjusted to 1 mL with 0.9% NaCl.
Macs multistand with macs separator
Manufactured by Miltenyi Biotec
The MACS MultiStand with µMACS separator is a lab equipment product from Miltenyi Biotec. It is designed to facilitate magnetic cell separation and sorting procedures. The core function of this product is to provide a stable and organized platform for the µMACS separator, enabling efficient and controlled magnetic separation of cells or other biological samples.
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Lab products found in correlation
2 protocols using macs multistand with macs separator
1
Immunoaffinity Isolation of Urinary Extracellular Vesicles
2
Isolation of Urinary Extracellular Vesicles
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Immunoaffinity isolation of uEVs was performed using a commercially available kit (Exosome Isolation Kit Pan, human, Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer's instruction. The kit is based on the immunomagnetic isolation of uEVs carrying any of the surface epitopes CD9, CD63 or CD81. Briefly, pre-processed void urine was precleared by centrifugation at 10.000 x g for 30 minutes (4°C). Then, 10 mL of the supernatant were concentrated using an Amicon Ultra-15 100 kDa filter (regenerated cellulose; Merck/Millipore) by centrifuging at 4.000 x g for 10 minutes. The concentrate was adjusted to 2 mL with 0.9% NaCl. Then, 50 µl of antibody-loaded magnetic beads were added. After incubation for 1 hour at room temperature the sample was loaded onto an equilibrated separation column within a magnetic stand (MACS MultiStand with µMACS separator, Miltenyi Biotec). After serial washing, uEV-bead-aggregates, the column was removed from the magnetic stand. uEV-bead aggregates were eluted by flushing with 100 µl isolation buffer. The samples were adjusted to 1 mL with 0.9% NaCl.
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