beta_gal_staining
ß-gal Whole-mount Skin
| Staining solution |
| 930 µl 1X Staining solution |
| 10 µl Staining supplement A |
| 10 µl Staining supplement B |
| 50 µl 20 mg/ml X-gal in DMF |
Samples are supplied in 4% PFA, rinse twice in PB for 5 minutes
Prepare 1X staining solution with 900 µl dH2O & 100 µl 10X staining solution
Prepare staining solution in PP tubes only!
Add 1 ml staining solution to sample, incubate at 37 ºC in humidifier overnight
Rinse the tissue with PBS , fix in 4% PFA overnight
Rinse the tissue with PBS twice for 10 minutes each time
Blot off PBS and dehydrate in 15 ml Sarstedt tubes on rocker table
Microtome (4 µm) on pink superfrost slides
Dry at 37 °C overnight
Dewax slides
Rinse in water and stain with Eosin Y (0.5% w/v in dH20 with 5% Acetic Acid) for one minute (dependant on staining)
Rinse in hot water, dry and mount with fluoromount
| Dehydration protocol |
| 70% ethanol | 2x 15’ |
| 98% ethanol | 2x 15’ |
| 99.5% ethanol | 2x 15’ |
| Isopropanol | 2x 15’ |
| Paraffin (56-58°C) | 3x 15’ |
| Dewaxing protocol |
| Isopropanol |
| 100% Ethanol |
| 95% Ethanol |
| 70% Ethanol |
| 50% Ethanol |
beta_gal_staining.txt · Last modified: 2021/01/19 11:10 by 127.0.0.1
