Chromatography Forum

Hi..
How much is a typical protein loaded on the analytical TSK gel 3000 SW XL SEC column 7.8*300 mm..
What is the column capacity?
Thanks!

Hi..
How much is a typical protein loaded on the analytical TSK gel 3000 SW XL SEC column 7.8*300 mm..
What is the column capacity?
Thanks!

This is a Size Exclusion Chromatography (SEC) column and it doesn't have a capacity for protein since it doesn't bind any protein. The protein are separated by molecular weight (size) through a kind of filtering process.
Typically, you would not load more sample than 2% of the total column volume. So, in this case, don't load more than 285 ul of your sample. The protein concentration of the sample is of less importance but keep it reasonable concentrated (>1 mg/ml).
Of course, it all depends on the complexity of your sample. How many proteins are you separating? How close in molecular weight are they? Are you looking for impurities or aggregation? etc...

Hi..
How much is a typical protein loaded on the analytical TSK gel 3000 SW XL SEC column 7.8*300 mm..
What is the column capacity?
Thanks!

This is a Size Exclusion Chromatography (SEC) column and it doesn't have a capacity for protein since it doesn't bind any protein. The protein are separated by molecular weight (size) through a kind of filtering process.
Typically, you would not load more sample than 2% of the total column volume. So, in this case, don't load more than 285 ul of your sample. The protein concentration of the sample is of less importance but keep it reasonable concentrated (>1 mg/ml).
Of course, it all depends on the complexity of your sample. How many proteins are you separating? How close in molecular weight are they? Are you looking for impurities or aggregation? etc...

thanks!
ya.. im trying to separate dimers and higher aggregates.. impurity testing..