with deionized water. Store at 4
C. Add 5 μL of Tween 20 to
obtain a final concentration of 0.005% if the running buffer is
going to be used on a SA sensor chip with biotinylated ligands
(see Note 2).
2. BIAcore extra wash solution: 50% isopropanol, 1 M NaCl, and
50 mM NaOH. Weigh 0.4 g of NaOH and 11.688 g of NaCl
and transfer to a glass beaker. Add about 100 mL of deionized
water to the glass beaker. Mix and transfer to a 100 mL
graduated cylinder and adjust to 100 mL with deionized
water to obtain a final concentration of 2 M NaCl and
100 mM NaOH. Mix this solution with an equal volume of
isopropanol.
3. BIAcore regeneration solution: 1 M NaCl and 50 mM NaOH.
Weigh 0.4 g of NaOH and 11.688 g of NaCl and transfer to a
glass beaker. Add about 200 mL of deionized water to the glass
beaker. Mix and transfer to a 200 mL graduated cylinder and
adjust to 200 mL with deionized water.
4. BIAcore detergent solution A: 1% β-octylglucoside. Weight
0.5 g of β-octylglucoside and transfer to a glass beaker. Add
50 mL of deionized water to the glass beaker. Mix and transfer
to a 50 mL graduated cylinder and adjust to 50 mL with
deionized water.
5. BIAcore detergent solution B: 0.5% sodium lauryl sulfate
(SDS). Weight 0.25 g of sodium lauryl sulfate and transfer to
a glass beaker. Add 50 mL of deionized water to the glass
beaker. Mix and transfer to a 50 mL graduated cylinder and
adjust to 50 mL with deionized water.
Fig. 6 A fixed concentration of analyte is injected with increasing concentrations
of peptide (a) or lipid (b) over immobilized peptide on a BIAcore SA sensor chip.
Note a competition with the peptide in solution (a) and a positive cooperative
effect with the lipid (b) over the immobilized peptide
PDZ-Peptide and PDZ-Lipid Interactions by SPR
79