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