How can we optimize the conditions for protein binding to nitrocellulose surfaces?
Normally the binding to the nitrocellulose membrane is very easy to achieve. However, in cases where the isoelectric point (pI) is only achieved at very low or high pHs, or if the proteins are very small, conditions may need to be fine tuned to improve or optimise binding to nitrocellulose surfaces. As several different forces (hydrophobic interactions, electrostatic interactions and hydrogen bonding) contribute to the binding between protein and nitrocellulose, the optimal conditions for proper binding, as well as maintaining protein activity (e.g. enzymes, antibodies), have to be carefully validated in biochemical assays. For optimal protein binding, the pH of the printing buffer should be close to isoelectric point of the printed protein. In order to receive stable attachment of the proteins, a drying step immediately after printing and before any additional processing steps (e.g. blocking), is strongly recommended. Different proteins show varying binding affinities to nitrocellulos
Normally the binding to the nitrocellulose membrane is very easy to achieve. However, in cases where the isoelectric point (pI) is only achieved at very low or high pHs, or if the proteins are very small, conditions may need to be fine tuned to improve or optimise binding to nitrocellulose surfaces. As several different forces (hydrophobic interactions, electrostatic interactions and hydrogen bonding) contribute to the binding between protein and nitrocellulose, the optimal conditions for proper binding, as well as maintaining protein activity (e.g. enzymes, antibodies), have to be carefully validated in biochemical assays. For optimal protein binding, the pH of the printing buffer should be close to isoelectric point of the printed protein. In order to receive stable attachment of the proteins, a drying step immediately after printing and before any additional processing steps (e.g. blocking), is strongly recommended. Different proteins show varying binding affinities to nitrocellulos
