i am performing SPR experiment and i am referring some journal and websites they stated regarding coupling buffer and immobilization buffer so ihave confusion about these buffer.
I think you could make a distinction between covalent attachment of the protein to the chip (coupling) and non-covalent attachment, such as by an affinity tag. Both are forms of immobilization. There may be requirements for a coupling buffer that do not apply to a non-covalent immobilization buffer because of the chemistry involved.
sir actually i am immobilizing my protein in CM5 chip. My protein present in tris , NaCl and ph 7.4. So can use these buffer in SPR.
For amine coupling on a carboxylated dextran chip (CM5) the buffer requirement is that it is free of amines. So PBS, HEPES are good buffers, TRIS not.
For more basic answers see www.sprppages.nl
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