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Differencial
Scanning Calorimeter
(VP-DSC, Malvern)
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Differential scanning
calorimetry (DSC) enables accurate
measurement of temperature
dependent properties. The
stability of a structure can be
investigated, the melting
temperature and the complete
thermodynamics of melting can be
determined.
Applications include the
measurement or characterisation of
- protein
stability and folding,
- the stability
nucleic acids, membranes, lipids
and micelles,
- the effects of
structural change on a molecule’s
stability,
- ultra-tight
interactions (up to 1020
M-1),
- other,
pharmaceutical parameters.
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Also for DSC, the solution
conditions can be readily altered to allow
determination of the pH or salt dependence
of thermodynamic quantities.
Alternatively, CD spectroscopy measures
the melting of structures using their
optical characteristics, and the Thermofluor
Assay might serve
as a much higher throughput method to
screen various conditions.
Sample Requirements
- Volume: 850 μl
- Concentrations: depends on the
protein, e.g. 0.1 mg/ml for lysozyme
- Protein needs to be quite pure. A
single band on an overloaded gel is a
good indication
Buffer Requirements
- Volume: 2 x 850 μl
- Avoid buffers with pKa
dependence on the temperature such as
Tris
- All reducing agents and particularly
DTT are causing baseline artefacts. A
test experiment is recommended
References:
Brandts
& Lin, Biochemistry 29, 6927-6940
(1990)
Gill
et al, J. Biomol. Tech. 41(4), 167-193
(2010)
Please
contact us for detailed protocols
and planning your experiment. |
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