Skip to main content

Table 2 Specifications for IHC in formalin-fixed and paraffin-embedded sheep brain tissue and Western/dot blot antibody dilution

From: Gene and protein patterns of potential prion-related markers in the central nervous system of clinical and preclinical infected sheep

    IHC Western/dot blot
Gene symbol Description Type/commercial reference Dilution epitope retrieval Visualization system Positive control Dilution
COL1A2 collagen 1 alpha 2 Mouse monoclonal against Collagen I Abcam AB6308 1:100 HIER Citrate buffer pH 6 10’ Pressure cooker Minimum 30’ cooling DAKO EnVision anti mouse Spleen (moderate perivascular and capsule staining, trabeculae unstained). 1:1500
COL3A1 collagen 3 alpha 1 Rabbit polyclonal against Collagen III Abcam AB7778 1:100 DAKO EnVision anti rabbit Spleen (moderate perivascular staining and mild staining of trabeculae). 1: 5000
GALA1 galanin 1 Rabbit polyclonal ENZO Life sciences BML-GA1161-0100 1:400 Brain tissue (stains diffusely the neuropile of cerebellar molecular layer) 1:1000
MTNR1B melatonin receptor 1b Rabbit polyclonal Sigma SAB29002 12 1:100 Retina (stains the plexiform layers). 1:1500
CAPN6 calpain 6 Rabbit polyclonal Abcam AB76974 1:50 HIER Citrate buffer pH 6 20’ water bath at 95ºC Minimum 30’ cooling DAKO EnVision anti rabbit Placenta (stains placental epithelium). 1:1000
MT2A methallothionein 2A Mouse monoclonal against MT1+2 Dakocytomation M00639 1:200 DAKO EnVision anti mouse Brain tissue (stains stellate shaped glial cells). 1:1500