To my dismay I have looked and looked trying to find the most reliable way to make a 1% Brilliant Blue G-250 solution as a stock to make other staining solution from. Among its physical properties is the insolubility (partial) in water and even in ethanol, which is mentioned across the available information. However, nobody out there dares to post a simple protocol explaining how to make a stable and reliable solution of the colloidal version of this dye. Does anyone have any solutions? I refuse to believe there isn't any out there.
Well, it is tricky. You should do the following:
1) Mix the Brillant blue with the water (for example 2 g of stain in 100 ml of water), stirring.
2) In between mix a 340 ml of Methanol + Phosphoric acid (35ml)+ 300ml of water
3) Add to the water/methanol/acid Ammonium sulfate slowly (wait for dissolve), and keep stirring until the salt will be completely dissolved (you should add water to reach 900 ml). After of that, keep stirring for 2 or 3 hours more.
4) Add the Water/brilliant blue to the methanol/water/acid/salt solution and keep stirring
5) Leave washout stirring overnight.
6) Filtered and use it
Dear Sharad, if I don't guess that you would like to stain Gels, I would like to point you to: http://cdn.preterhuman.net/texts/science_and_technology/nature_and_biology/Genetics/Proteomics%20in%20Practice%20-%20A%20Laboratory%20Manual%20of%20Proteome%20Analysis%20-%20Tom%20Naven.pdf
Reiner Westermeier, Tom Naven (Eds):
Proteomics in Practice: A laboratory manual of Proteome Analysis
© 2002 WILEY-VCH Verlag GmbH,
ISBN’s 3-527-30354-5 Hardcover 3-527-60017-5 (electronic)
go to page “259 of 329” (right margin display position to turn –drag down pages rapidly):
= real book page 233
find: STEP 4: Staining of the Gels
1 Colloidal Commassie Brilliant Blue Staining
Recipe for Stock solution A & B and how to proceed: on p.233 & 234
Hopefully this hint could be of help too solving your problem...Best regards and good luck
Hello again after a long period(!) of stagnation in this thread:
a) Thanks to Md Sanower Hossain (:-));
b) it would be interesting to see whether the original requester has any opinion now ("However, nobody out there dares to post a simple protocol explaining how to make a stable and reliable solution of the colloidal version of this dye. Does anyone have any solutions? I refuse to believe there isn't any out there").
c) two other possibilities out of my files (NB:cave (C) Copyright! Find two pdf's enclosed (see next RE-post)
best regards,
Wolfgang
Two other possibilities out of my files (NB:cave (C) Copyright!)
(no.1)
Regards,
Wolfgang
Two other possibilities out of my files (NB:cave (C) Copyright!)
(no.2)
Regards,
Wolfgang
Since some minute ago I found another posting thread concerning the matter, I would like to add:
"Regarding protein re-staining"
I have done Coomassie Brilliant Blue (CBB) staining for protein detection then destained the gel for removal of excess dye not bound to proteins.
Now I want to remove the dye bound to proteins to go for silver staining with same gel. But in the destaining solution the dye bound to proteins will not remove completely. Could you suggest any other opinions?
Mar 28, 2012 • Modified Apr 1, 2012 by the author
to be found @
http://www.researchgate.net/topic/Gel_Electrophoresis/post/Regarding_protein_staining
which also should point anybody in need to the TOPIC GelElectrophoresis @
http://www.researchgate.net/topic/Gel_Electrophoresis/
Regards, Wolfgang
Have you tried adding glacial acetic acid to the solution at 10% concentration V/V? Stirring overnight using a magnetic stirer should do the job then.
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining