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Immunohistochemical reactions were performed using SignalStain Boost IHC Detection Reagent and SignalStain DAB Substrates Kit (both from Cell Signaling Technology Inc., Danvers, MA, USA).
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DAB and Vector Blue Substrate Kit (Vector Laboratories) were used for enzymatic substrate.
Bound primary Abs were detected using AmpliStain™ HRP conjugate (SDT GmbH, Baesweiler, Germany) with NovaRed substrate kit.
HRP labelling was visualised using a NovaRed substrate kit (Vector Laboratories, Burlingame, CA, USA).
Bound primary Abs were visualized using AmpliStain™ HRP conjugate (SDT GmbH, Baesweiler, Germany) with NovaRed substrate kit.
Horseradish peroxidase-conjugated secondary antibodies and an enhanced chemiluminescence substrate kit were used for the detection of specific proteins.
Reactions were visualised by the Opti-4CNTM substrate kit (Bio-Rad) used according to manufacturer's instructions.
HRP was reacted with DAB (Peroxidase Substrate Kit; Vector), and alkaline phosphatase with Fast Blue/Napthol AS-MX (Sigma-Aldrich).
Membranes were washed and signals were revealed with the SuperSignal WestPico Chemiluminescent Substrate Kit (Thermo Scientific Pierce, Waltham, MA, USA).
Biotin-SP-conjugated Biotin-SP-conjugated Goat AntiRat IgG+IgM H+Lry antibody from JacksecondaryoReseantibodyoratories (West Grove, PA) and Vectastain ABC kit and DAB Peroxidase Substrate kit from Vector Laboratories (Burlingame, CA) were used.
Bound primary antibody was detected using a biotinylated anti-sheep IgG secondary antibody, ABC reagent (Vector) and DAB Substrate kit (Sigma-Aldrich) according to the manufacturers' instructions.
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