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