Solubilising peptides - (Aug/16/2007 )
I need to dissolve my lyophilised BSA-conjugated peptide for use in an ELISA. The data sheet says: sparingly soluble in water: Use DMSO to dissolve prior to adding water. I ended up having to use a lot of DMSO, and even now I am not sure whether it has completely dissolved. Would the presence of DMSO affect the ELISA? Does anybody have any suggestions for dissolving peptide?
Thank you.
-suds-
QUOTE (suds @ Aug 16 2007, 09:04 PM)
I need to dissolve my lyophilised BSA-conjugated peptide for use in an ELISA. The data sheet says: sparingly soluble in water: Use DMSO to dissolve prior to adding water. I ended up having to use a lot of DMSO, and even now I am not sure whether it has completely dissolved. Would the presence of DMSO affect the ELISA? Does anybody have any suggestions for dissolving peptide?
Thank you.
Thank you.
In which part of the ELISA are you using the peptide/BSA? Why do you think the peptide might not have dissolved completely? How much peptide did you get, and how much DMSO have you added (that is, what concentration is the peptide?)?
-swanny-
QUOTE (swanny @ Aug 17 2007, 01:29 AM)
QUOTE (suds @ Aug 16 2007, 09:04 PM)
I need to dissolve my lyophilised BSA-conjugated peptide for use in an ELISA. The data sheet says: sparingly soluble in water: Use DMSO to dissolve prior to adding water. I ended up having to use a lot of DMSO, and even now I am not sure whether it has completely dissolved. Would the presence of DMSO affect the ELISA? Does anybody have any suggestions for dissolving peptide?
Thank you.
Thank you.
In which part of the ELISA are you using the peptide/BSA? Why do you think the peptide might not have dissolved completely? How much peptide did you get, and how much DMSO have you added (that is, what concentration is the peptide?)?
I assume you will be coating the peptides to your ELISA plate. I should imagine that almost all of the DMSO would be removed when you conduct your washing after the coating incubation has been completed. It would not be a bad idea to have some sort of control... perhaps a nonsense/irrelevant peptide resuspended and coated in the same conditions?
-hpylori-
QUOTE (hpylori @ Aug 17 2007, 11:46 AM)
QUOTE (swanny @ Aug 17 2007, 01:29 AM)
QUOTE (suds @ Aug 16 2007, 09:04 PM)
I need to dissolve my lyophilised BSA-conjugated peptide for use in an ELISA. The data sheet says: sparingly soluble in water: Use DMSO to dissolve prior to adding water. I ended up having to use a lot of DMSO, and even now I am not sure whether it has completely dissolved. Would the presence of DMSO affect the ELISA? Does anybody have any suggestions for dissolving peptide?
Thank you.
Thank you.
In which part of the ELISA are you using the peptide/BSA? Why do you think the peptide might not have dissolved completely? How much peptide did you get, and how much DMSO have you added (that is, what concentration is the peptide?)?
I assume you will be coating the peptides to your ELISA plate. I should imagine that almost all of the DMSO would be removed when you conduct your washing after the coating incubation has been completed. It would not be a bad idea to have some sort of control... perhaps a nonsense/irrelevant peptide resuspended and coated in the same conditions?
Hyplori, you are correct. I do have a scrambled peptide to use as a control. Thank you.
-suds-