I need protocol of stain for SDS-page for small amount of proteins (1-2 ug per band).
Dear Hanna,
1-2 ug protein is easily visualized with standard proteins stain such as Coomassie Blue Stain; you can purchase it as G-250 or R250 powder and make up your own solution or purchase the already made up solution.
For protein amounts less than ~25 ng, you can advance to a more sensitive dye, "Simply Blue", aka "GelCode Blue" stain. This is still Coomassie based, easy to use stain. Silver stain is more sensitive than the Coomassie based stains; it can help visualize protein amounts in low ng amounts.
Good luck,
Hediye
Dear Hediye,
Thank you for your answer. I used Coomassie Blue Stain (G250), but in protocol says that minimum amount for this stain is 20 ug...And i can not found proteins in the band (I have 1-2 ug)...Can you give me references for this stain with 1-2 ug proten, please?
And can you give me protocol for "Simply Blue" stain, please?
Best regards,
Hanna
Dear Hannah,
I think that there has to be an error in the protocol, mixing ug with ng; if you do a quick search, in Researchgate or in peer reviewed publications, you will agree that limit of detection of Coomassie Blue stain is about 100ng. My own experience confirms that the limit of detection is probably close to 30ng of protein. Is it possible that the protocol you have means 20ng instead of 20ug?
The colloidal Coomassie stain, "Simply Blue" is sold by Invitrogen (through Life Technologies):
https://www.lifetechnologies.com/order/catalog/product/LC6060
You can request it from the supplier as a sample to try - I have never made it up from powder..
Best, Hediye.
Dear Hediye,
I think that there was no error, since This protocol gave me in the lab, where it is used for a long time, and besides, I have to electrophoresis to determine the concentration of protein by the Bradford and initially took a different concentration - a concentration of less than 20 ug I no trace of protein staining with Coomassie, etc. I have not found. Perhaps when stained by Coomassie have any micromethod and I used makrometod?
Best regards,
Hanna
Use silver stain some proteins are not well stained with comassie blue.
Thank you! I used silver stain, but this staining does not reflect the amount of protein. It shows only the presence of protein...
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