Antibodies

2.2.1. Staining Antibodies

1. Anti-CD3e, clone M-20 (Santa Cruz Biotechnology).

2. Anti-CD3^, carboxy terminal binding antibody, clone C-20 (Santa Cruz Biotechnology).

3. Anti-FcRy (Upstate Biotechnology, Waltham, MA).

4. Anti-linker for activated T cells (LAT), clone-FL-233 (Santa Cruz Biotechnology).

5. Antiactin, clone-I-19 (Santa Cruz Biotechnology).

2.2.2. Isotype Controls

1. Goat immunoglobulin G (IgG) (Santa Cruz Biotechnology).

2. Rabbit IgG (Santa Cruz Biotechnology).

3. Mouse IgG (Santa Cruz Biotechnology).

2.2.3. Blocking Antibodies

Human IgG (Sigma Aldrich, St Louis, MO) was used for blocking antibodies.

2.2.4. Secondary Fluorescent-Labeled Antibodies

All the immunoglobulins have minimum crossreactivity with bovine, chicken, goat, guinea pig, Syrian hamster, horse, human, mouse (except three), rat, and sheep IgG.

1. Tetramethylrhodamine isothiocyanate (TRITC)-labeled donkey antirabbit IgG (H + L) (Jackson ImmunoResearch Lab, West Grove, PA).

2. TRITC-labeled donkey antigoat IgG (H + L) (Jackson ImmunoResearch Lab).

3. Fluorescein isothiocyanate (FITC)-labeled antimouse IgG (H + L) (Jackson ImmunoResearch Lab).

4. FITC-labeled donkey antirabbit IgG (H + L) (Jackson ImmunoResearch Lab).

2.2.5. Secondary HRP-Labeled Antibodies

1. Goat antirabbit IgG-HRP (Santa Cruz Biotechnology).

2. Antigoat IgG-HRP (Santa Cruz Biotechnology).

3. Antimouse IgG-HRP (Santa Cruz Biotechnology).

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