From: Lymphocytic infiltration in the cutaneous lymphoma microenvironment after injection of TG1042
Antibody | Expression | Clone | Isotype | Reference | Source | Antigen retrieval | Dilution | Amplification system |
---|---|---|---|---|---|---|---|---|
CD3 | T-lymphocytes | rabbit pc IgG | A0452 | DAKO | EDTA buffer pH9 (Target Retrieval®, Dako) | 1/100 | 0.5% dilution of horse biotinylated secondary antibody + ABC amplification system, Vector Laboratories | |
45 minutes - 95°C | ||||||||
CD4 | helper and regulatory T-lymphocytes | 1 F6 | mouse mc IgG1 | NCL-CD4-1 F6 | NOVOCASTRA | 10 mM Citrate Buffer pH6 | 1/20 | non diluted anti-mouse Envision® system, Dako |
4B12 | mouse mc IgG1 κ | NCL-CD4-368 | NOVOCASTRA | 3 x 5 minutes – 750 W | 1/20 | non diluted anti-mouse Envision® system, Dako | ||
CD8 | cytotoxic T-lymphocytes | C8/144B | mouse mc IgG1κ | #M7103 | DAKO | EDTA buffer pH9 (Target Retrieval®, Dako) | 1/50 | non diluted anti-mouse Envision® system, Dako |
45 minutes - 95°C | ||||||||
CD20cy | mature and immature B-lymphocytes | L26 | mouse mc IgG2a κ | M7055 | DAKO | 10 mM Citrate Buffer pH6 | 1/200 | 0.5% dilution of horse biotinylated secondary antibody + ABC amplification system, Vector Laboratories |
2 minutes - 750 W + 7 minutes -250 W | ||||||||
CD56 | Natural Killer cells | 564 | mouse mc IgG2b | NCL-CD56-504 | NOVOCASTRA | 10 mM Citrate Buffer pH6 | 1/50 | non diluted anti-mouse Envision® system, Dako |
45 minutes – 95°C | ||||||||
CD79αcy | B cells from early pre-B to plasma cell stage and neoplastic B-lymphocytes | HM57 | mouse mc IgG1κ | M7051 | DAKO | EDTA buffer pH9 (Target Retrieval®, Dako) | 1/50 | 0.5% dilution of horse biotinylated secondary antibody + ABC amplification system, Vector Laboratories |
45 minutes - 95°C | ||||||||
TIA-1 | CD4 and CD8 positive T cells, granulocytes, monocytes and activated NK cells | 2G9 | mouse mc IgG1 | PN IM2550 | IMMUNOTECH | 10 mM Citrate Buffer pH6 | 1/50 | non diluted anti-mouse Envision® system, Dako |
2 minutes - 750 W + 7 minutes -250 W | ||||||||
MHC Class I | most nucleated cells | HC10 | mouse monoclonal IgG2a | SB03 546 | kindly provided by Dr S. Ferrone, G. Gaslini Institute, Italy | EDTA buffer pH9 (Target Retrieval®, Dako) | 1/20 | non diluted anti-mouse Envision® system, Dako |
45 minutes - 95°C. |