Can you Destain Coomassie with water?
Staining with Coomassie Blue R250 Destain the gel by soaking for at least 2 hours in 10% acetic acid, 50% methanol, and 40% H2O with at least two changes of this solvent. If the gel still has a Coomassie Blue background then continue destaining until the background is nearly clear.
What is the fastest way to Destain Coomassie gel?
FAST DESTAINING OF COOMASSIE GELS. Extract the gel from the sealing system (BioRad) alter running and introduce into a little tupperware with aprox 50ml of Coomassie solution (AcH : MeOH : H2O 10:45:45 with 0.25% Brilliant Blue G-250). Shake at RT and change de destaining solution until the bands can be observed.
What is Coomassie blue staining used for?
Description. Coomassie blue dyes are a family of dyes commonly used to stain proteins in SDS-PAGE gels. The gels are soaked in dye, and excess stain is then eluted with a solvent (“destaining”). This treatment allows the visualization of proteins as blue bands on a clear background.
How do you make a Destain solution?
Destain: Add 500mL of HPLC- grade methanol to 300 mL of HPLC grade water. Add 100 mL of reagent grade acetic acid and, after mixing, adjust the total volume to 1000mL with water. The final concentrations are 50% (v/v) methanol in water with 10% (v/v) acetic acid.
How do you Destain instant blue?
Protocol for Gel Drying : Processing the gel prior to full incorporation of stain may result in destaining and reduced sensitivity, this can be reversed by incubating gel in InstantBlue overnight. 2) Submerse the gel in approximately 100 ml ultrapure water at ~70°C (heat for 30s to 60s in a microwave oven).
Can you reuse Coomassie stain?
The Coomassie stain is removed by decanting. You may reuse the stain so pour it into a new vial. Cover the gel with the destain solution and allow the gel to destain with gentle agitation. The destain solution should be changed several times, removing it at each change by aspiration.
How long should I stain my gel?
Stain the gel at room temperature for 3 to 4 hr with gentle agitation. The Coomassie stain is removed by aspiration after staining. 4. Cover the gel with ~250mL of the destain solution and allow the gel to destain with gentle agitation.
What does the G 250 in the Coomassie Brilliant Blue G 250 mean?
The suffix “R” in the name of Coomassie brilliant blue R-250 is an abbreviation for Red as the blue colour of the dye has a slight reddish tint. For the “G” variant the blue colour has a more greenish tint. The “250” originally denoted the purity of the dye.
What is the detection limit of Coomassie Blue staining?
0.3 to 1 ug/
The detection of protein bands in the gel by Coomassie Blue staining depends on nonspecific binding of a dye, Coomassie Brilliant Blue R, to proteins. The detection limit is 0.3 to 1 ug/protein band.
Can you Destain overnight?
If you destain overnight, you can also decrease the time in destain so that your bands are still dark when the background has destained sufficiently. You can also restain with GelCode Blue if you are using that instead of a traditional coommassie stain. Bands are faint because the protein loaded are very few.