Anti-CD3 & CD28 antibobdy nanoparticles
|Cat No.||Size||Price||Add Cart|
Anti-CD3 & CD28 antibobdy nanoparticles are ideal to be used to activate human T cells. The nanoparticles are conjugated with both of anti-human CD3 antibody and anti-human CD28 antibody. They act as antigen-presenting cells and activate resting T cells from peripheral blood mononuclear cells (PBMCs) as well as purified T cells.
Anti-CD3 & CD28 antibody nanoparticles are provided in phosphate buffered saline (PBS) with 0.01% sodium azide. 1 ml of suspansion solution is enough for expansion of 4X107 human T cells. Store at 4°C up to 6 months.
Additional material required
Human T cell activation/expansion
For optimal results from the nanoparticles, it is recommended that the nanoparticles are washed prior to addition to samples.
1. Vortex MagVig nanoparticles for 10-20 seconds.
2. Transfer the desired volume of nanoparticles to a tube. Add equal volume of Buffer, and vortex to mix.
3. Separate the nanoparticles from the solution by placing the magnet on the side of the tube for 2-5 min and remove the supernatant carefully (with magnet still on the side). Note: A clear precipitate containing dark brown colored nanoparticles should become visible on the side of the microcentrifuge tube.
4. Remove the tube from the magnet and resuspend the washed nanoparticles in the same volume of Culture Medium as the initial volume taken to wash.
Activation of Human T Cells
1. Add 25 ul of nanoparticles to 1 ml cells (1X106/ml).
2. Incubate in a humidified CO2 incubator at 37°C, according to your specific requirements.
3. Harvest the activated T cells and use directly for further analysis.
4. For flow cytometry applications, remove nanoparticles prior to staining. Place the tube on a magnet for 2-5 min to separate the nanoparticles from the solution. Transfer the supernatant containing the cells to a new tube.
Expansion of Human T Cells
1. Add 25 ul of SI-nanoparticles to 1 ml cells (1X106/ml).
2. Add 30 U/ml rIL-2.
3. Incubate in a humidified CO2 incubator at 37°C, according to your specific requirements.
4. Check cells daily. Add rIL-2 every other day. When the cell density exceeds 2.5 X 106 cells/ml or medium turns yellow, split cultures to a density of 0.5-1 X 106 cells/ml in culture medium containing 30U/ml rIL-2.
5. Count cells on day 5, 7, 9 and 11.
6. For human pan-T cells, recommend to culture cells for 11 days.
Table 1. Guild for the use of bead volumes.
|Seeding T cell numbers||8X104||1X106||50X106|
|bead volume||2 ul||25 ul||1.25 ml|
|rIL-2||30 U/ml||30 U/ml||30 U/ml|
|seeding medium volume||100-200 ul||1-2 ml||50-100 ml|