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Table 2 Antibodies used for immunohistochemical evaluation

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.  
  1. MHC Class I: Major Histocompatibility Complex I (Human Leukocyte Antigen A, B, C); mc: monoclonal; pc: polyclonal; TIA-1: T-cell restricted intracellular antigen-1.