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Table 2 Summary of immunohistochemical methodology

From: Activation of regulated cell death in the lung of piglets infected with virulent PRRSV-1 Lena strain occurs earlier and mediated by cleaved Caspase-8

Specificity Type of antibody Commercial origin Blocking solution Dilution Antigen retrieval
TUNEL N.A In Situ Cell Death Detection Kit, POD, Roche, Germany N.A N.A Proteinase K in heat incubator
cCasp3 (Asp175) mAb SignalStain Apoptosis Cleaved Caspase-3 IHC Detection Kit, Cell Signaling, USA 5% NGS 1:500 pH 6 citrate buffer in microwave
PRRSV (SDOW 17) mAb Rural Technologies, Brookings, SD, USA 2% BSA 1:500 Protease XIV in water bath
CD163* (2A10/11) mAb INIA, Madrid, Spain 2% BSA Neat pH 3.2 citrate buffer microwave
Arginase-1 pAb ThermoFisher Scientific, Barcelona, Spain 2% BSA 1:100 pH 8.5 citrate buffer microwave
cCasp8 (Asp391) mAb Cell Signaling; Danvers, MA, USA 2% BSA 1:100 Proteinase K in heat incubator
cCasp9 (Asp330) mAb Cell Signaling; Danvers, MA, USA 2% BSA 1:50 Proteinase K in heat incubator
  1. mAb: monoclonal antibody, pAb: polyclonal antibody, N.A.: not applicable, NGS: Normal goat serum, BSA: Bovine serum albumin (Sigma-Aldrich, Taufkirchen, Germany); Proteinase K (Roche, Basel, Switzerland), 15 min at 37 °C in heat incubator; Protease Type XIV (Sigma-Aldrich), 8 min at 37 °C in water bath. *CD163 mAb was kindly provided by Dr. J. Domínguez, INIA, Madrid, Spain.