BioSciences - Polysciences, Inc.

304
Additional Particles
Protein Binding Protocols
Microspheres & Particles
The following sequences serve as guidelines for protein binding. Technical Data Sheets (TDS) with detailed step-by-step protocols can
be downloaded from our website.
Adsorbing Protein on Particles
TDS #238E
Plain Polystyrene
• Initial Buffer: 0.1M Borate Buffer,
pH 8.5
• Suspend in buffer, spin down and
resuspend 2 or 3 times.
• Suspend in borate buffer.
• Add protein and mix end-to-end
overnight.
• Spin and save supernatant for
protein determination.
• Resuspend in BSA in appropriate
buffer and spin down twice.
• Resuspend in PBS, pH 7.4,
containing BSA and glycerol
(storage buffer).
• Protein bound directly on surface.
Coupling by Carbodiimide
TDS #238C & #644
Carboxylate Functional Particles
• Initial Buffer: 0.1M Carbonate Buffer
• Suspend in buffer, spin down and
resuspend 2 or 3 times.
• Suspend in phosphate buffer.
• Add fresh carbodiimide solution
dropwise and incubate for 15-30
minutes.
• Wash to remove excess
carbodiimide, then resuspend
in borate buffer.
• Add protein and mix end-to-end
for 2-4 hours.
• Add ethanolamine, mix for
30 minutes.
• Spin and save supernatant for
protein determination.
• Resuspend in BSA-containing
buffer and spin down twice.
• Resuspend in PBS, pH 7.4,
containing BSA and glycerol
(storage buffer).
• Protein bound 2-3 carbon atoms
from surface.
For more information please call (800) 523-2575 or visit: polysciences.com
Coupling by Glutaraldehyde
TDS #238D & #238G
Amino or Blue Dyed Particles
• Initial Buffer: 0.02M PBS, pH 7.4
• Suspend in buffer, spin down and
resuspend 2 or 3 times.
• Suspend in PBS.
• Suspend in 8% glutaraldehyde in
PBS, pH 7.4, and mix end-to-end
for 4-6 hours.
• Wash to remove excess
glutaraldehyde and resuspend in
PBS buffer.
• Add protein and mix end-to-end
overnight.
• Add ethanolamine, mix for 30
minutes.
• Spin and save supernatant for
protein determination.
• Resuspend in BSA-containing
buffer and spin down twice.
• Resuspend in PBS, pH 7.4,
containing BSA and glycerol
(storage buffer).
• Protein bound 5 carbon atoms
from surface of blue dyed beads
and 11-12 carbon atoms from
surface of amino beads.