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Author Topic: ESSENTIAL PRECAUTIONS TO BE NOTED WHILE WORKING WITH PROTEINS  (Read 6123 times)
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Francis Umeoguaju
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« on: April 10, 2010, 08:26:31 am »


Some basic precaution to be exercised while working with protein are listed below.

Protein extracts must always be kept in solutions with pH that are near the physiological pH where the protein is found naturally or at the pH that gives optimum activity for the protein.

Protein extracts must be kept in a buffered media that would resist a change in the Ph of the protein solution since change in pH can lead to denaturation and precipitation of the protein.
   
Care should be taken when deciding which type of buffer to use with the protein extract since the constituent of some buffer alter or inhibit the activity of the enzyme or may interface with the method of assay of the enzyme. For instance a phosphate buffer will not be used for an enzyme that require calcium co-factor since the calcium will form precipitate. Also a Tris buffer will not be used for an enzymatic reaction that would be assayed with Folin-Lowry method since the Tris would give false result with folin reagents.
   
The protein extract or pure protein samples should be kept under refrigeration as this would stop any proteolytic activity of proteases in the crude extracts. It will also prevent microbial degradation of the extract.
   
The media and apparatus to be used for the extraction and purification process such as acetone, mortal and pestle etc. should be refrigerated as this would reduce the inherent denaturation ability of these extraction media on the extracts.
   
When preparing a protein solution, the protein should be dissolved gently with a glass rod and should not shaken vigorously as would denature the protein and give erroneous result.
   
All extraction process should be carried out at a temperature of about 0-5 degree C  to reduce protein denaturation.
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