1.

Protein Denaturation
a. Denaturation causes by acetic acid addition

5 mL protein solution

Entered into test tube

Added 2 drops acetic acid 1N while shaking
Flake (protein precipitate)

b. Denaturation
Heated incauses
waterby heating
bath in 5 minutes
Observed the changes in precipitation
2-3 mL
Observation of protein solution
result
Entered into test tube
Heated until 1 minute
Turbid solution (White flake)

Cooled down
Distributed

Part 1 Part 2

Added 1-2 drops of Heated

(NH4)2SO4
(ammonium sulfate) Protein precipitation
Heated
c. Denaturation causes by formaldehyde addition
Protein
1-1,5 precipitation
ml formaldehyde + 2 mL aquades

Entered into test tube

Added protein solution drop by drop
Observation result

2. Amfoter properties of protein

a. Amfoter properties of protein (from egg) testing

3 mL of aquadest

Entered into test tube

Added 1 drop HCl 1N
Added some drops of congo indicator
Blue solution

Added 2-3 mL protein solution

b. Test with Note
alkaline
theatmosphere
color changes

Color changes
 Test tube I Test tube II
3 mL of NaOH 0,1 M 2-3 mL protein solution

Entered into test tube Entered into test tube

Added a few drops of the PP Added drops NaOH 0,1 M
indicator Observed the color changes that
Pink solution occur

Test tube III Color changes

Test tube IV
3 mL of dilute NaOH 2-3 mL protein solution

Entered into test tube Entered into test tube

Added a few drops of the PP Added drops NaOH 0,1 M
indicator Observed the color changes that
Pink solution occur
Color changes

3. Precipitation of Protein
a. Precipitation of Protein with ammonium sulphate
3-4 mL of protein solution

1. Put in test tube

2. Added 3-4 mL saturated ammonium sulphate
3. Shake slowly
TheTurbid
4. solution becometaken
solution turbid1 mL
5. Transfered to another test tube
6. Added 2-3 mL aquades
7. Shake it

b. Deposition of Protein with mineral acids

Dissolve
Test tubeprecipitate
I (colorless)

1 mL of HNO3 concentrated

Entered into test tube

Test tube filted
Added 1-1,5 mL drop by drops of protein solution
through the tube wall
The test tube is re-established
White ring formed as a protein precipitate

Shake again
Added 1 mL HNO3 concentrated
More precipitate
 Test tube II
1 mL of HCl concentrated

Entered into test tube

Test tube filted
Added 1-1,5 mL drop by drops of protein solution
through the tube wall
The test tube is re-established
White ring formed as a protein precipitate
c. Deposition of Protein with heavy metals
Shake again
Added -2 mL HCl concentrated
1-1,5 mL protein solution 1-1,5 mL protein solution
More precipitate
Poured into test tube Poured into test tube
Added CuSO4 drop by drop Added PbSO4 drop by drop
while shaking it while shaking it
Blue precipitate Blue precipitate

1-1,5Added CuSO4
mL protein 0,1 M
solution 1-1,5Added PbSO4
mL protein 0,1 M
solution
Dissolve
Pouredprecipitate
into test tube Dissolve
Pouredprecipitate
into test tube
Added ZnSO4 drop by drop Added FeSO4 drop by drop
while shaking it while shaking it
Blue precipitate Blue precipitate

4. Protein3. Added
color ZnSO4 0,1 M
reaction 3. Added FeSO4 0,1 M

Dissolve
a. Biuret precipitate
reaction Dissolve precipitate

3 mL protein solution

Entered into test tube

Added 1 mL 40% NaOH
Added drop by drop 0,5% CuSO4, the color become red
or purple
b. Chantoprotein reaction
Red or purple solution
3 mL protein solution
Put in a test tube
Added 1 mL of concentrated HNO3
Heated the solution mixture

Yellow solution
Cooled
3 mL protein solution
Added
c. Ninhydrin ammonia drop by drop
reaction
Set the pH until pH = 7
Orange
Takensolution
1 mL
Put in a test tube
Added 10 drops of ninhydrin 0,2%
Heated the solution mixture at 100˚C for 10 minutes
Observed the color changes that occur
Observation Result
 d. Millon reaction
2 mL protein solution
Put in a test tube
Added 1 mL of reagent Millon
Heated the solution mixture

Yellow solution
Cooled with water
Added 1 drop of 1% NaNO2 solution
Reheated precipitate or solution
e. Hopke-Cole reaction
Orange solution
3 mL protein solution
Put in a test tube
Added 1 drop of dilute formaldehyde
Added 1 drop of mercuriculphate reagent
Added 1 mL of concentrated H2SO4 through the filted
tube wall
Heated the solution mixture at 100˚C for 10 minutes
Observed the color changes that occur
Formed 2 layers
5. Protein hydrolysis (the for
and test upper field shows a purple ring and if it’s
sulfur
shaken the whole solution become purple)
1 mL protein solution

Put in a test tube

Added 1 mL of 40% NaOH solution
Heat the mixture solution for 1 minute
Added 1 drop of Pb-acetate

Black precipitate (PBs)