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Classification of Proteins
Conjugated
Simple Proteins Derived Proteins
Proteins
1- Solubility of proteins
Identify the solubility of different types of proteins in different solvents.
Principle:
Simple proteins dissolve easily in water while soluble protein complex by low
solubility in distilled water. The proteins are derived insoluble in water but
soluble in alkaline solutions.
How to perform the test:
1. Test the solubility of each (albumin, casein, gelatin, peptone) in cold water
and then in hot water and then in sodium hydroxide solution.
2. Record solubility of all proteins in cold water and hot water and sodium
hydroxide solution in the following table of results.
2-Precipitation of proteins
(A) Precipitation of proteins by heavy metals:
Principle:
Proteins are precipitated in alkaline medium with heavy metals due to the
direct union of cation (Cu++, Ag+, Ba++, Pb++) with anionic groups of proteins,
which are formed in basic medium.
At alkaline pH 7 and above, proteins are usually negatively charged so the
addition of positively charged ions will neutralize this charge and the proteins
come out of solution (i.e. heavy metals combine with proteins forming
insoluble metalloproteine).
How to perform the test:
1- Put 2ml from protein solution (albumin, casein, gelatin, and peptone) in
4 test tubes.
2- Add 3 drops from silver nitrate (AgNO3). Or other heavy metals are
found in lab.
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3-Color Tests
(A) Biuret Test:
Principle:
It is the general test for all proteins. Biuret reagent is dilute CuSO4 in strong
alkaline medium. Alkaline CuSO4 reacts with all compounds containing 2 or
more peptide bonds to give a blue-violet color.
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5- FreezingTest:
Principle:
In this test we see the change shape at low temperatrue due to change shape,
chemical and physical properties of proteins.
How to perform the test:
1-Put 3ml from protein solution (albumin, casein, gelatin, and peptone) in 4
test tubes.
2- Put the tube in the Cup by the snow and leave for five minutes.
3- Notice formation of freezing of the solution (give with gelatin only).
4- Record your result in the table.
6- Sulfur test:
Principle:
The basic idea of this test is to detect the presence of amino acids that contain
sulfur such as cystein.
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7- Phosphorus Test
Principle:
The basic idea of this test is to detect the presence of amino acids that contain
How to perform the test:
1-Put 3ml from protein solution (albumin, casein, gelatin, and peptone) in 4
test tubes.
2 - Add 3 mL of alkaline solution of 20% NaOH.
3 - Heat in a boiling- water bath for 10 minutes.
4-Add in the order as follows: (1 ml of Molbidic acid and + 1 ml sodium
sulphate + 1 ml of 0.5% hydroquinone ) respectively.
5- Formed blue colour (gives with casein only).
6-Record your result in the table.
8-Millon's test:
Principle:
The basic idea of this test is to detect the presence of amino acid by the
hydroxyl group in proteins such as phenyl acid tyrosine.
How to perform the test:
1-Put 1ml from protein solution (albumin, casein, gelatin, and peptone) in 4
test tubes.
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9- Hopkin-cole Test
Principle:
The basic idea of this test is to detect the presence of amino acids that contain
indol group such as tryptophan.
How to perform the test:
1-Put 3ml from protein solution (albumin, casein, gelatin, and peptone) in 4
test tubes.
2 - Add 3 mL of Hopkin-cole reagent.
3 – Mix well.
4-Add carfuley conc H2SO4 down side of tube so to form two layers.
5- Formed purple ring colour.
6-Record your result in the table.
When you are finished collecting data, determine the identity of each
unknown. Record chemical properties of your test, observation and results in
the tables below.
Unknown (1)
Unknown (2)
Unknown (3)
Unknown (4)