Dear researchers,
May I know do anyone can suggest the protocol of protein elution from SDS PAGE ?
Any simple and inexpensive methods?
Thank you.
Hi Jennifer
You should have been more specific with your question... It isn't clear what u actually want to do with the protein.
However, i am attaching some protocols that might help you out.
Good luck
What do you mean with elution? And what would you want to do afterwards with your protein?
Hi Jennifer,
What you want is to take out a protein directly from the gel or from a nitrocellulose or PVDF membrane, once it has been transferred, for example in order to sequence it?
hi Jennifer
what do you mean ? you must be more specification in your qusition ?
Hi Jennifer
You should have been more specific with your question... It isn't clear what u actually want to do with the protein.
However, i am attaching some protocols that might help you out.
Good luck
Dear researchers,
I would like to describe my problem in details.
I would like to do elution of protein after carrying out separation of protein by SDS-PAGE.
After getting the bands from the SDS-PAGE, I have to cut out the bands and do the elution before sending them to LC-MS.
So, I would like to know how to do the elution of protein? May I know what is the steps to do this elution?
Anyone can suggest any simple and inexpensive methods of elution?
Thank you.
You can cut the band from the gel with and put it in an eppendorff with few microliters of the loading buffer for electrophoresis. You can then chop the piece of gel up, vortex it and also let it in contact with the buffer for a while fir sonication which helps as well as the protein band can pass into the buffer. Then just spin the eppy down and recover the buffer with your protein. You can than concentrate it, dialyse it or reload it directly on another gel if you want to run it again.
Good luck!
Junaid Furhan
Thanks for your explanation.
But, I would like to ask about the article (TR0051) which you suggest to me.
The part {elute the protein from the gel matrix}
step 2:
elution buffer (50 mM Tris-HCl, 150 mM NaCl, and 0.1 mM EDTA; pH 7.5)
May I know do you have this recipe? Because I don't know the exact amount of needed to add.
For 1L preparation
Add 50 ml of 1M Tris HCl, 37.5 ml of 4 M NaCl and 33.62 mg EDTA.
ddH2O to 1L
Can I replace ddH20 with dH20?
Because My university dont have ddH20...
Junaid Furhan
- Badges
- Science method