Determining Protein Melting Points

Micromeritics’ NanoPlus nano particle size and zeta potential analyzer comes with a temperature gradient feature to determine the size and zeta potential of proteins. With the help of this exclusive feature, researchers can further use the integrated Peltier thermo-electric controller for temperature analyses.

Sample Cell

The NanoPlus analyzer features a sample cell that can be held at specific temperatures ranging from 5°C to 90°C or a gradient of decreasing or increasing temperatures in increments of 0.1°C or above. This is perfect for determination of either the melting or denaturing point of proteins using the NanoPlus.

Temperature Gradient Setting

In order to set up a temperature gradient for protein analysis, the ‘Manual Temperatures Setting’ under ‘Measurement Parameters’ should be set to ‘Gradient’ in the Size Standard Operating Procedures (SOP) Designer. Next, the ‘Gradient Temperature (°C)’ table featuring the end temperature, start temperature, interval size and equilibration time must be completed. Through this setting, the NanoPlus is prompted to obtain size data at all temperature points and save each temperature point as an individual data file in the ‘Size Analysis’ database.

After the analysis has been completed, the software will automatically produce a ‘Temperature Gradient Analysis’ file, which can be easily opened under ‘Size Analysis’. This file will end as .tgr for temperature gradient and will show Size (nm) versus Temperature (°C) when it is opened. The cumulative values obtained will be utilized for the size points.

Melting point of BSA was found to be at 68.0°C.

Figure 1. Melting point of BSA was found to be at 68.0°C.

In this example a sample of bovine albumin serum (BSA) was tested by means of the temperature gradient function. The test was carried out twice and in both instances the melting point was observed to be at 68.0°C temperature.

Conclusion

The unique temperature gradient feature in the NanoPlus instrument makes it possible to determine the melting/denaturing points of proteins. Additionally, the Peltier Thermo-electric controller incorporated in the NanoPlus allows for temperature analyses.

This information has been sourced, reviewed and adapted from materials provided by Micromeritics Instrument Corporation.

For more information on this source, please visit Micromeritics Instrument Corporation.

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