Comassie Brilliaant Blue staining takes so much time
If anyone have tried other methods with reliable results
many thanks
Coomassie Blue staining can be sped up by heating the gel + stain in the microwave. Destaining can also be sped up this way.
You can also use colloidal Coomassie Blue stains that require no destaining step. InstantBLue from Expedeon allows staining in 15 minutes without destaining.
http://www.expedeon.com/instantblue-protein-stain.html/
I've actually heard some older researchers swear by radioactive labeling as faster and more reliable, although I can't attest to it from first-hand experience.
Coomassie Blue staining can be sped up by heating the gel + stain in the microwave. Destaining can also be sped up this way.
You can also use colloidal Coomassie Blue stains that require no destaining step. InstantBLue from Expedeon allows staining in 15 minutes without destaining.
http://www.expedeon.com/instantblue-protein-stain.html/
Keep the gel in water just sufficient to dip the gel in water add 400-500 microlit of chloroform shake it for 5-7 mins. Wash with water. visualize the gel on UV . U have to wait for some 4-5 mins to band to appear. Sensitivity as good as comassie.
You can use trichloro-acetic acid. Either add it to the gel and running buffers, or use it as a one step stain/fix solution. You can then image the proteins using a UV light box (or the gel doc system used for DNA gels).
Use CBB-G 250 instead of R-250. Use Bradford reagent which contains G-250 for staining. First heat the gel in microwave for 2 mins and keep it on shaker for 5 mins. Then add Bradford reagent and boil. You will see bands. No need to destain. If you don't have microwave use hot water.
You can add 2,2,2 trichloroethanol to your gel solution before polymerization (50 mkl to 10 ml) and then after running a gel visualize by UV. Actually this will be a home made stain-free visualization - you can use stain-free ode at BioRad geldocs. But sensitivity of the CBB is much better.
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