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Welcome to the Integrated Department of Immunology at the University of Colorado - School of Medicine and National Jewish Health.

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Biacore System Setup

*Biacore should have been left in H2O


 
  1. Run DESORB (under working tools menu):  This should be done ~1/week.  Removes protein stuck in system, should be run with the maintenance chip in the BIACORE.
    •    3ml Buffer 1 in R2F3 – stored in drawer in Biacore room
    •    3ml Buffer 2 in R2F4 – stored in KM lab cold room
  2. Run SANITIZE (working tools):  This should be done once a month. Should also be run with the maintenance chip docked.
    •    Mix 1.5ml Bleach (Biacore brand) w/ 20ml H2O in the supplied 50ml Falcon tube.
    •    Come back before leach is done running to put H20 back on
  3. Dock New Chip
    •    UNDOCK maintenance chip (command menu)
    •    DOCK experimental chip (command menu) 
    *If chip has been run before then skip to step 4
    •    Replace H20 with HBST buffer
    •    NORMALIZE (working tools) with 40% glycerol in position R2F2
    •    FLUSH  (working tools) w/HBST to flush out glycerol
  4. PRIME/FLUSH (working tools) system w/ HBST
  5. Run amazing Nobel Prize worthy experiments
  6. When finished replace the maintenance chip, change buffer to H2O.
  7. If leaving for 4 days or less leave it on STANDBY (working tools) with 100ml of H2O.
  8. If leaving for longer do not leave it running.
 
 
 
HBST running buffer
5ml 1M TC grades HEPES (10mM)
15ml 5M NaCl (150mM)
3.4ml .5M EDTA (3mM)
250μl 10% Tween 20 (0.005%)        Pierce Cat# 28320 (Surfact Amp 20)
Bring up to 500ml in ddH2O, filter and degas
 
 
 
Biacore Solutions for covalently linking peptides to a CM5 chip
EDC
Make up 100 μl aliquots of 0.4M EDC in H2O and store at –20
 
NHS
Make up 100 ul aliquots of 0.1M NHS in H2O and store at-20
 
TCEP-HCl
Make up 100 μl aliquots of 1M TCEP-HCl and store at –20.
 
Also need, 0.1M NaOH, 1M EthyleneDiamine, 40% glycerol.