Mike Barnkob:Protocols/Immunology/Making tetramer: Difference between revisions
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# Calculate: Volume of Streptavidin = volume of monomer x 1.2 <br />'''Note:''' For 50μL monomer = 60 μL streptavidin. | # Calculate: Volume of Streptavidin = volume of monomer x 1.2 <br />'''Note:''' For 50μL monomer = 60 μL streptavidin. | ||
# Adjust concentration of both monomer and streptavidin to 1 mg / mL. | # Adjust concentration of both monomer and streptavidin to 1 mg / mL. | ||
# Add 1/3 of streptavidin volume to monomer and leave at 4°C O/N or 2 hours. Keep in the dark. | # Add 1/3 of streptavidin volume to monomer and leave at 4°C O/N or 2 hours. Keep in the dark.<br />'''Note:''' Carefully add the streptavidin. Do not pipet much up and down, and do not vortex. | ||
# Add 1/10 of streptavidin volume to monomer every 20 minutes. Keep at 4°C and in the dark. | # Add 1/10 of streptavidin volume to monomer every 20 minutes. Keep at 4°C and in the dark. | ||
===References=== | ===References=== | ||
Revision as of 08:40, 20 July 2015
Making Tetramers
Protocol for conjugating monomers to streptavidin-APC.
Reagents and equipment
- Monomers
- Straptavidin-APC (or other streptavidin-flurochrome conjugate)
Protocol
- Calculate: Volume of Streptavidin = volume of monomer x 1.2
Note: For 50μL monomer = 60 μL streptavidin. - Adjust concentration of both monomer and streptavidin to 1 mg / mL.
- Add 1/3 of streptavidin volume to monomer and leave at 4°C O/N or 2 hours. Keep in the dark.
Note: Carefully add the streptavidin. Do not pipet much up and down, and do not vortex. - Add 1/10 of streptavidin volume to monomer every 20 minutes. Keep at 4°C and in the dark.
