|88179S||1 Kit (50 tests)||
|Product Includes||Quantity (with Count)||Storage Temp|
|Rapid-Act Anti-human CD3 Mouse mAb||1 x 125 µg||+4°C|
|Rapid-Act Anti-human CD28 Mouse mAb||1 x 125 µg||+4°C|
|Rapid-Act Synthetic Immune Synapse||1 x 500 µl||+4°C|
Additional Reagents (Not Supplied)
NOTE: Volumes are suitable for stimulation and expansion of 1 x 106 cells. For different cell numbers, adjust volumes according to Table 1.
NOTE: We recommend preparing the Rapid-Act CD3/CD28 complex fresh for each experiment. If necessary, store pre-mixed components at 4 °C. Do not freeze.
Table 1: Recommended volumes for different cell numbers
|Cell number||5 – 8 x 104||2 – 5 x 105||0.5 – 1 x 106|
|Cell culture medium (ml)||0.1 – 0.2||0.5 – 1.0||1.0 – 2.0|
|Rapid-Act CD3/CD28 complex (µl)||3||15||30|
1. T cell activation
2. T cell expansion
NOTE: Activation markers CD25 and CD69 should be upregulated after 48 h.
Protocol Id: 2786
T cells are activated by signaling through ITAM (immunoreceptor tyrosine-based activation motif)-containing CD3 signaling chains that associate with the T cell receptor (1). Co-stimulation through CD28 on T cells provides an additional signal required for effective T cell activation (2). Antibody-based binding and aggregation of CD3 and CD28 proteins simulates the engagement of a T cell with an antigen presenting cell, and results in T cell activation.
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