LAB report Cell biology 20.

11
SDS - Polyacrylamide Gel Electrophoresis (SDS-Page)
Methodes and Materials

Team D4
Anna Siebert

Materials 1
- Electrophoresis chambers
- SDS-Page gels
- Cell extract from MiaPaCl2
- Protein sample
- Protein ladder
- Running buffer
- Staining buffer (10% acetic acid | 25% methanol | 0.05% Coomassie Brilliant Blue G-
250)
- Destaining buffer (10% acetic acid | 30% methanol | 60% distilled H2O)

Methods
1. Set up the electrophoresis apparatus.
2. Ad buffer solution to each of the three samples X (unknown protein, concentration x), Y
(also unknown protein, concentration y) and Z (cell extract). Afterwards heat the
samples in a water bath for approximately fiveteen minutes at 70C.
3. Load 10μl of each sample on the prepared gel.
4. Run electrophoresis at 120V for one hour until the dye is nearly at the bottom of the
gel.
5. Stain the gel with staining buffer on a rocking table for about fiveteen minutes.
6. Rinse the gel afterwards two times for ten minutes with destaining buffer (also on a
rocking table).
7. Leave it in deionised water bath overnight to enlighten.
8. On the next day determine the protein by comparing it to the protein ladder.

1
https://rockland-inc.com/SDS-Page-Electrophoresis.aspx (28.11.2019)