Guidelines to prepare SDS samples
Prepare samples for SDS page gel
This section aims to give a guideline to run a commercial 15 combs SDS page gel.
For the SDS page gel you will need 4 tubes
| Sample | Lab Grade Water | 3.6x Loading reducing dye | |
|---|---|---|---|
| Eluted Protein | 3 µL | 5.7 µL | 3.3 µL |
| Core + NFW (negative control) | 3 µL | 19 µL | 9 µL |
| Crude CFPS | 3 µL | 19 µL | 9 µL |
Standards
To have a semiquantitative assay, you can prepare and run BSA standards alongside your samples.
Dilute your BSA sample to 1 mg/mL and make dilution as outlined in the table below to prepare your 3 standards (A, B and C)
| Required Stocl | Lab Grade Water | 3.6x Loading reducing dye | |
|---|---|---|---|
| Standard A | BSA 1 mg/mL | 24 µL | 176 µL |
| Standard B | Standard A | 50 µL | 150 µL |
| Standard C | Standard B | 50 µL | 150 µL |
Load the SDS PAGE gel
Load on the stain-free protein gel, 4-15% (15 well):
- 4 µL pre-stained protein ladder
- 4 µL eluted scaled-up protein sample
- 8 µL eluted scaled-up protein sample
- 4 µL No-DNA negative expression control
- 4 µL positive expression control
- 4 µL BSA standard A
- 4 µL BSA standard B
- 4 µL BSA standard C
Run at 200 V for 40 minutes