Home >
Community >
What is the role of glycine in the running buffer for SDS-polyacrylamide gel electrophoresis
Upvote
30
Downvote
+ Biochemistry
Posted by
Mike Jonas
What is the role of glycine in the running buffer for SDS-polyacrylamide gel electrophoresis
The pH of stacking and resolving gel are set in such a way that one is above and one is below the pI of gly (5.97).Therefore, Gly has two different charges in stacking and resolving gel. That is in stacking gel gly has no charge (which will allow the protein to stack according to size in this part of the gel rather than separating because of presence of gly in between the protein molecules, constraint in separation of proteins) but in resolving gel charge is negative since the pH is higher (constraint in movement of proteins no longer present so separation of protein based on size possible).
The pH of stacking and resolving gel are set in such a way that one is above and one is below the pI of gly (5.97).Therefore, Gly has two different charges in stacking and resolving gel. That is in stacking gel gly has no charge (which will allow the protein to stack according to size in this part of the gel rather than separating because of presence of gly in between the protein molecules, constraint in separation of proteins) but in resolving gel charge is negative since the pH is higher (constraint in movement of proteins no longer present so separation of protein based on size possible).
Could you add a reference or link to allow for background reading?More
Upvote
VOTE
Downvote
since the stacking gel have a ph of 6.8 the glycine will attain a neutral charge(by the isoelectric point and ph relation)thus the chloride ions travel faster followed by the sample and then at the last glycine ions,thereby stacking the sample in between both.when it reaches the resolving gel the ph increases which gives glycine a negative charge.so they travel faster along with the chloride ions leaving back the sample to get seperated without any hinderence
since the stacking gel have a ph of 6.8 the glycine will attain a neutral charge(by the isoelectric point and ph relation)thus the chloride ions travel faster followed by the sample and then at the last glycine ions,thereby stacking the sample in between both.when it reaches the resolving gel the ph increases which gives glycine a negative charge.so they travel faster along with the chloride ions leaving back the sample to get seperated without any hinderence
The pH of stacking and resolving gel are set in such a way that one is above and one is below the pI of gly (5.97).Therefore, Gly has two different charges in stacking and resolving gel. That is in stacking gel gly has no charge (which will allow the protein to stack according to size in this part of the gel rather than separating because of presence of gly in between the protein molecules, constraint in separation of proteins) but in resolving gel charge is negative since the pH is higher (constraint in movement of proteins no longer present so separation of protein based on size possible).
Hope this was helpful..
The pH of stacking and resolving gel are set in such a way that one is above and one is below the pI of gly (5.97).Therefore, Gly has two different charges in stacking and resolving gel. That is in stacking gel gly has no charge (which will allow the protein to stack according to size in this part of the gel rather than separating because of presence of gly in between the protein molecules, constraint in separation of proteins) but in resolving gel charge is negative since the pH is higher (constraint in movement of proteins no longer present so separation of protein based on size possible).
Hope this was helpful..
More
VOTE
VOTE
since the stacking gel have a ph of 6.8 the glycine will attain a neutral charge(by the isoelectric point and ph relation)thus the chloride ions travel faster followed by the sample and then at the last glycine ions,thereby stacking the sample in between both.when it reaches the resolving gel the ph increases which gives glycine a negative charge.so they travel faster along with the chloride ions leaving back the sample to get seperated without any hinderence
since the stacking gel have a ph of 6.8 the glycine will attain a neutral charge(by the isoelectric point and ph relation)thus the chloride ions travel faster followed by the sample and then at the last glycine ions,thereby stacking the sample in between both.when it reaches the resolving gel the ph increases which gives glycine a negative charge.so they travel faster along with the chloride ions leaving back the sample to get seperated without any hinderence
More
VOTE
VOTE