In general, Coomassie Brilliant Blue (CBB) is used for protein staining after SDS-PAGE. This staining shows high background and needs hours to overnight de-staining to get clearly visible staining bands. CBB Rapid Stain is a product that gives you visible bands within one hour after SDS-PAGE. In addition, longer de-staining gives you visible bands a few ng of proteins.
Features:
1. 30 ng protein band visible within 60 min
2. A few ng protein band visible by extending staining and de-staining
3. Highly clear bands can be seen by extending de-staining
4. Long storage stability due to 2 liquids formulation
General quick method
This method is intended to detect around 30 ng of protein bands within one hour. The following is an example using one mini size gel.
1) Wash: After SDS-PAGE, wash the gel with 50 mL of DW for 10 min using a shaker. Wash twice.
2) Staining: Immerse the gel into 20 mL of Solution A, add 2 mL of Solution B, and mix the solutions through roughly. The gel staining is started. Use enough volume of solutions so that the gel is totally soaked. Bands can be seen 5min after start of staining. Normally 30 min is enough to detect 30 ng of protein bands.
3) Wash: Wash the gel with DW to remove excess staining solution.
4) Storage: Store stained gel in DW. Stained bands become more clearer while storing due to destaining.
5) De-staining (optional): To save time de-staining can be done by using 30 ml of 1% acetic acid. The de-staining time has to be determined by observing bands (normally 15 min). Store in DW.
High sensitivity method
This method is intended to detect a few ng of protein bands or to get more clearer bands. Additional fixing and de-staining solutions (both recipes described below) are needed. Following is an example using one mini size gel.
1) Fixation: After SDS-PAGE, fix the gel with 30mL of fixing solution (50% MeOH+10% Acetic acid) for 10 min using a shaker.
2) Wash: wash the gel with 50 mL of DW for 10min.using a shaker. Wash twice.
3) Staining: Immerse the gel into 20 mL of Solution A, add 2 mL of Solution B, and mix the solutions though roughly. Use enough volume of solutions so that the gel is totally soaked. The gel staining is started. Stain for 1 to several hours while shaking. The staining time has to be determined by observing bands.
4) Wash: Wash the gel with DW to remove excess staining solution.
5) De-staining: Soak the gel in 30 ml of de-staining solution (1% acetic acid). The de-staining time has to be determined by observing bands (normally 15 min). Change de-staining solution to accelerate the de-staining.
