Direct Detect® Spectrometer User Guide
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8
General Guidelines
The Direct Detect® spectrometer is recommended for use with protein samples ranging from 0.25 to
5 mg/mL. Although the instrument can measure protein concentrations from 0.1 mg/mL to around
30 mg/mL, the most accurate results are achieved within the recommended concentration range. The
instrument is also capable of detecting and analyzing lipids and detergents. Analysis is performed with
a user-defined calibration method. Samples to be analyzed are introduced to the system using the
Assay-free card which contains a novel IR-compatible membrane. The recommended sample volume is
2 µL per spot position.
8.1 Buffer components
The Direct Detect® system is compatible with most commonly used buffers, detergents, and
reducing agents, however, use of buffers which contain amides or amine functional groups
may affect quantification.
Buffers containing more than 5% glycerol, SDS, and/or Tween® 20 surfactant should be
avoided due to the excessive time required to achieve dryness.
The instrument comes with preloaded protein calibration files, prepared using National
Institute of Standards & Technology (NIST) bovine serum albumin (BSA) in phosphate-
buffered saline (PBS). However, preparation of a fresh calibration method with the
buffer used for protein preparation is recommended. The Direct Detect® spectrometer is
compatible with most solvents used in lipid research.
8.2 Materials Required but Not Supplied
● Protein standards (Bovine Serum Albumin - National Institute of Standards &
Technology SRM 927d (optional))
● Protein sample(s)
● Lipid/detergent standards
● Lipid/detergent sample(s)
● Fixed-volume precision pipette capable of precisely dispensing 2 µL, and pipette tips
NOTE:
To minimize pipetting error, we recommend the use of a fixed-volume precision
pipette capable of dispensing 2 µL, for applying samples to the Assay-free card.
Adjustable pipettes are not recommended.
● Water, PBS, or other buffer
● Chloroform or other lipid compatible solvent
● Printer (optional)
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