MaxSortin™CD4 Isolation Kit

The MaxSortin™ CD4 Separation Kit can be used to sort human CD4+ T cells. Through incubating nanoscale CD4 separation magnetic beads with cells, the sorting of CD4+ T cells can be achieved. The nanoscale CD4 separation magnetic beads are incubated with peripheral blood mononuclear cells (PBMCs) and then subjected to magnetic separation, which enables the separation and enrichment of CD4+ T cells, playing a role in purifying CD4+ T cells and is applicable to the production of cell therapy products.

1.1 Resuspend human PBMCs in PBS buffer containing 1% human serum albumin (HSA). Take a sample for counting and transfer 1×10⁷ cells into a 1.5 mL EP tube. Then centrifuge at 1500 rpm for 5 minutes.
1.2 Discard the supernatant, take 80 μL of MaxSortin™ Cell Sorting Buffer to resuspend the cells, add 20 μL of CD4 separation magnetic beads, mix well and then place them in a refrigerator at 2 - 8 °C for incubation for 15 minutes.
1.3 Take the MaxSortin™ L-Separation Column and place it on the magnetic-activated cell sorting (MACS) sorter, and rinse it twice with 1 mL of MaxSortin™ Cell Sorting Buffer.
1.4 Take the sample that has completed incubation out of the refrigerator at 2 - 8 °C, add 1 mL of MACS solution, centrifuge at 1500 rpm for 5 minutes, and discard the supernatant.
1.5 Add 1 mL of MaxSortin™ Cell Sorting Buffer to resuspend the cells, add the sample into the separation column. After it flows out naturally, add MaxSortin™ Cell Sorting Buffer in two portions, 1 mL each time. Collect the effluent in a 15 mL centrifuge tube.
1.6 After all the MaxSortin™ Cell Sorting Buffer has flowed out, remove the separation column from the MACS sorter and place it in another new 15 mL centrifuge tube. Add 3 mL of MaxSortin™ Cell Sorting Buffer into the separation column and use the piston that comes with the separation column to push the liquid out directly.
1.7 Place the 15 mL centrifuge tube containing the collected liquid into a horizontal centrifuge and centrifuge at 1500 rpm for 5 minutes.
1.8 After centrifugation, pour out the supernatant, take 1 mL of 1× Dulbecco's phosphate-buffered saline (DPBS) solution to resuspend the cells, count the cells and conduct flow cytometry detection.