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BIAcore/Biosensor Analysis Request Form

Key information about the BIAcore Technology:

Macromolecular interactions are visualized in real time with unlabelled ligands and analytes.

Surface plasmon resonance (SPR) senses changes in refractive index of the accumulation of molecules on the gold sensorchip surface.  Responses are continuously recorded and plotted against time (sensorgrams).

Data can be obtained on antigen-antibody interactions, isotyping, epitope mapping, DNA-protein interactions and various other macromolecular complex formation or cleavage reactions. Association/dissociation rate constants are determined from sensorgram data.

Ligand coupling to the sensorchip is accomplished by amine, ligand or surface thiol, and aldehyde covalent bonds as well as by affinity-capturing via biotin/avidin, antibodies, histidine tags, and liposomes.  Several modified sensorchips are available for a variety of requirements.  Ligands bound to the sensorchip can be regenerated multiple times to test a battery of analyte samples.

Sample Preparation Requirement:

Ligands for immobilization should be of 90% purity and dissolved in buffers that do not contain amines, Tris, BSA, glycine etc).  They must not contain salts >200 mM or substances of high refractive index such as glycerol or sucrose.  The ligand concentration should be 25 - 250 µg/ml, (consumption per immobilization ~ 10 - 100 µl).  M.W. should be > 5 kD.  Smaller ligand molecules can be captured and immobilized, although they will not produce an SPR signal without a reaction with a larger analyte.

Analytes should be greater than 5kD, contain no substances with high refractive index, and a concentration of 10 - 100 µg/ml, (consumption per test 20 - 50 µl).

Buffers should have salt concentrations of 150-200 mM and contain low amounts of detergent (0.005% P-20 or Tween-20). HEPES buffered saline with EDTA, pH 7.4, is preferred.  SDS, organic solvents or chaotropic reagents destroy the sensorchip. (For more information and references:  http://www.biacore.com)

Name of Principal Investigator:

 

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Division/Laboratory:

 

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Lab location: ________________ Phone: _____________ e-mail: ____________________________

Please attached a brief description of the goals of the experiment, concentrations and M.W. of ligand and analyte, buffers, etc:

 

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FUNDING & BILLING INFORMATION:

HRI ACCOUNT #: ________________    STATE ACCOUNT #: _______________
Please contact core personnel about the charge fee for usage.
PI will be billed quarterly.

APPROVED BY (ALL SIGNATURES ARE REQUIRED BEFORE USAGE)

PI: ____________________________________________ DATE: _________
LAB CHIEF: _________________________________DATE: _________
CORE DIRECTOR: ______________________________ DATE: _________

Print & Send completed form to:

Contact Personnel: Jane Kasten-Jolly, Ph.D. -- BIGGS/C500 473-8660
Core Director, Dr. David Lawrence BIGGS, C539; Phone 402-5684; Fax 474-1412