MaxSortin® CD3 Isolation Kit-T&L

Composition

Product Details

• Reactive Species: Human

• Magnetic Bead Endotoxin: <2 EU/mL

• Magnetic Bead Appearance: Brown liquid

Intended Use

The MaxSortin® CD3 Separation Kit can be used to sort human CD3+ T cells. Through incubating nanoscale CD3 separation magnetic beads with cells, the sorting of CD3+ cells can be achieved.

Experimental Procedures

1. Resuspend human peripheral blood mononuclear cells (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.

2. Take 80 μL of MaxSortin® Cell Sorting Buffer to resuspend the cells, add 20 μL of CD3 separation magnetic beads, mix well and then place them in a refrigerator at 2 - 8 °C for incubation for 15 minutes.

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.

4. Take the sample that has completed incubation out of the refrigerator at 2 - 8 °C, add 1 mL of MaxSortin® Cell Sorting Buffer, centrifuge at 1500 rpm for 5 minutes, and discard the supernatant.

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.

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.

7. Place the 15 mL centrifuge tube containing the collected liquid into a horizontal centrifuge and centrifuge at 1500 rpm for 5 minutes.

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.

Precautions

1. When incubating the magnetic beads with cells, it is necessary to mix them thoroughly to improve the sorting efficiency.

2. When using this product to remove and sort CD3+ cells, it is necessary to appropriately increase the amount of magnetic beads used.

3. The buffer and separation column included in this kit can be used for the initial experiment. If more experimental operations are involved, please purchase MaxSortin® Cell Sorting Buffer (Article Number: MS-BF100) and MaxSortin® L-Separation Column (Article Number: MS-CL01) separately.

Important Note

This product is only applicable to in vitro cell culture and cannot be directly used for clinical treatment.