Concentration of sample is 10-12 mg/ml and I am planning to use 4 ml DEAE cellulose resin. I need to know the amount of protein sample required to be loaded on the column to get good resolution?
Hi Ravinder,
By mentioning 4ml resin you probably meant the presowllen beads (1:1 slurry) packed into a column which you are going to use and I assume you are not doing any batch preparation. For column prep the binding capacities vary between 450-600 mg protein /100g DEAE cellulose.For 1:1 packed slurry that comes 4.5-6.0 mg protein/g sowllen beads.
Since you are planning to use 4ml packed slurry (~4g of sowllen beads), it should ideally bind 18-24 mg of protein. Since you are planning to load 10-12mg/ml, to stay in the safe side I would suggest reducing the protein concentration up to half, ie 5-6mg/ml or increasing the bed volume to double i.e. ~8.0 ml.
For further reading you can consider the attached file.
Hope that helps.
Hi Ravinder,
By mentioning 4ml resin you probably meant the presowllen beads (1:1 slurry) packed into a column which you are going to use and I assume you are not doing any batch preparation. For column prep the binding capacities vary between 450-600 mg protein /100g DEAE cellulose.For 1:1 packed slurry that comes 4.5-6.0 mg protein/g sowllen beads.
Since you are planning to use 4ml packed slurry (~4g of sowllen beads), it should ideally bind 18-24 mg of protein. Since you are planning to load 10-12mg/ml, to stay in the safe side I would suggest reducing the protein concentration up to half, ie 5-6mg/ml or increasing the bed volume to double i.e. ~8.0 ml.
For further reading you can consider the attached file.
Hope that helps.
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