Immunofluorescence staining protocol for co-staining of fetuin-A and GFAP in older human autopsy tissue via Tyramide Signal Amplification v1

Abstract

This staining was performed to detect fetuin-A in astrocytes. Fetuin-A and astrocytes were detected in paraffin sections (1 μm thickness) of formalin-fixed human brain tissue. Glial Fibrillary Acid Antigen (GFAP) was marked by a polyclonal rabbit-anti-human antibody (Spring Bioscience Cat# E18320, RRID:AB_1661177, dilution 1:50) and a polyclonal goat-anti-rabbit Alexa Fluor® 488 conjugated secondary antibody (Thermo Fisher Scientific Cat# A-11070, RRID:AB_2534114, dilution 1:300).Fetuin-A was detected by using a monoclonal IgG2a mouse-anti-human antibody (clone MAHS-1, dilution 1.0 µg/mL), raised against purified human fetuin-A in our laboratories. Antibody binding was detected by tyramide signal amplification using a secondary biotinylated polyclonal goat-anti-mouse antibody (