Specification

Reactive Species	Human
Endotoxin	<2EU/mL
Appearance	Brown liquid

Intended Use

ActCel T Cell Activation Reagent is synthesized by coupling a nanomatrix with anti-human CD3 antibody and anti-human CD28 antibody, and it can gently and efficiently activate T cells or PBMCs (Peripheral Blood Mononuclear Cells).

Experimental Procedures

1.1 Conduct the experiment at a titer of 1:200. The specific addition amounts are shown in the following table:

Culture Plate	Maximum Culture Volume	Total T Cell Number	Volume of T Cell Activation Reagent Added per Well
96-well	0.2 mL	0.3×10⁶	1 μL
48-well	1 mL	1×10⁶	5 μL
24-well	2 mL	2×10⁶	10 μL
12-well	4 mL	4×10⁶	20 μL
6-well	5 mL	5×10⁶	25 μL

1.2 Taking 1×10⁶ T cells as an example: Add 995 μL of X-VIVOTM 15 medium and 5 μL of T cell activation reagent (containing 200 IU/mL IL-2) to 1×10⁶ T cells. Then culture them at 37 °C under 5% CO₂ for 2 days.

1.3 After culturing for 2 - 3 days, centrifuge at 300g for 10 minutes, remove the supernatant, and add 2 mL of fresh complete medium to remove the T cell activation reagent. Then continue culturing at 37 °C under 5% CO₂.

1.4 Every 2 days, pipette the cell suspension thoroughly to mix evenly, add 2 times the volume of fresh complete medium (the ratio of the original volume to the supplementary volume is 1:2), and count the cells to calculate the amplification factor. Then culture them in a 37 °C, 5% CO₂ incubator. For example, if the medium is supplemented on day 4, and the original volume is 2 mL, 4 mL of medium should be added, and the final total volume will be 6 mL.

Precautions

When incubating the T cell activation reagent with cells, it is necessary to mix them thoroughly to improve the activation efficiency.
The titer of the T cell activation reagent can be adjusted within the range of 1:100 - 300.
The time for centrifuging to remove the T cell activation reagent can be optimized according to customer requirements.

Documents

ActCel T Cell Activation Reagent

Additional Precautions

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