Proteomic Report A-1-1 Adrian Jayadi (07120100005)

ABSTRACT
Proteomics is a study of proteins in order to give a comprehensive view of
structure and functions to understand the biological process

since they are the

functioning units. Determining the concentration of protein can be conducted by

using the Bradford test and read the absorbance using the spectrophotometer at 560
nm. Our result has shown that the concentration of sample A (50.01 mg/ml) is nearly
normal, and normal for sample B (60,92 mg/ml) compared to the normal range at 5590 mg/ml. The result of sample A can be happened due to errors in the working
method or lack of protein in the blood.
After determining the concentration, we determine the molecular mass of the
protein and the relative abundance of major protein and also the distribution of
protein among fractions by using the SDS-PAGE method. Larger molecule will travel
slower than the smaller molecules. There are two fractions which are Ep and Es. EP
fraction has more protein than the Es fraction. The quantification and determination
of single protein ( AFP ) can be more specificaly done by using the ELISA test.
ELISA test are used to detect the substances that have antigenic properties, that
primarily protein. The test involves the use of monoclonal antibodies and enzyme
linked antibodies. Our group result of te AFP concentration is normal compared to
the normal range that is <20 ng/ml in healthy person. By using O-Toluidine reagent,
blood sugar level are determind colorimetrically. The Absorbance is read at 630nm.
The blood glucose level of our reslut on sample A is 93.46 mg/dl and on sample B is
86.7 mg/dl. The result is still within the normal range that is between 70 110 mg/dl.

INTRODUCTION
Protein is a large molecule composed of one or more chains of amino acids in
a specific order determined by the base sequence of nucleotides in the DNA coding
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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

for protein. Protein is made from a long chain of 20 types of amino acids, it is linked
by the covalent peptide bond between the carboxyl group and amino group. Proteins
are required for the structure, function and regulation of the bodys cells, tissues and
organs. Each protein has unique functions. Proteins are essential components of
muscle, skin, bones and the body as a whole. Protein also functions as enxyme,
structural support, hormones, immune defence, etc. Protein are a synthesise from
gene through the transcription and tranlation process. Protein is also used for energy
sources in the body. There are four structures of protein which are :
1. Primary Structure
The primary structure of peptides and proteins refer to the linear number
and order of the amino acids present.
2. Secondary Structure
In the secondary structure of protein, the ordered array of amino acids in a
protein confer regular conformational forms upon that protein. There are
two main types of secondary structure which are alpha helix ( composed of
a single linear array of helically disposed amino acids) and beta sheet
( composed of 2 or more different regions of stretches of at least 5 10
amino acids ).
3. Tertiary Structure
Refers to the complete 3 dimentional structure of the polypeptide units.
There are interactions between the side chains ( R-groups ) of the various
amino acids. The bonds formed in the tertiary stucture is include the
hidrogen bond, salt bridges, disulfide bonds.

4. Quaternary Structure
The structure formed by monomer monomer interaction in an oligometric
protein. It is the overall protein structure that results from the agregation of
these polypeptides subunits.
Human serum is devided in to two major group of protein which are albumin
and globolin. Albumin and globulin are filling out about 96 % of total serum proteins.
The normal amount of total serum protein in a healthy adult is about 55 90 mg/ml.
Total Serum protein can be interpreted as the total amount of protein that is found in
the blood.
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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

Bradford test is one of several simple methods commonly used to determine
the total protein concentration of a sample. The method is based on the proportional
binding of the dye Coomassie to proteins. The more protein present, the more
coomassie binds. Furthermore, the assay is colorimetric ; as the protein
concentration increases, the color of the test sample beome darker. As the
coomassie preferentially binds to select amino acids and changes from cationic (+)
state to an anionic (-) state, its bound condition is best detected at the maximal
absorbance spectrum at 595 nm. The protein concentration of a test sample is
determined by the comparison to that of a series protein standards known to
reproducibly exhibit a linear absorbance profile in this assay. The protein standard
that is used in this Bradford test is the most widely used protein as the standard
Bovine Serum Albumin (BSA) to determine the human serum protein.
SDS-Page

stands

for

sodium

dodecyl

sulfate-polyacrylamide

gel

electrophoresis. The SDS portion is an anionic detergent, meaning that dissolved its
molecules have a net negative charge within a wide PH range. The SDS has a
hydrophobic tail that interacts strongly with protein chains. Each SDS molecule
contributes two negative charges, overwhelming any charge the protein ay have.
SDS binds to all proteins and breaks up the weak

( non covalent bonds ) of the

proteins, as such the protein will form one straight line. Protein separation by SDSPAGE can be used to estimate relative molecular mass, to determine the relative
abundance of major protein in a sample, and to determine the distribution of proteins
among fractions. There are two fractions which are Ep and Es. Ep fraction has more
protein than the Es fraction.
The purpose of ethanol precipitation is to reduce the solubility of protein.
Ethanol precipitation is one method for removing SDS and other alcohol soluble
impurities from protein samples with minimal protein loss. The protein lysate was
precipitated with ethanol and subjected to SDS PAGE with quantitative Coomassie
blue staining.
Glucose is our body main source of energy. This type of sugar comes from
digesing carbohydrates into chemical that we can easily convert to energy. We get
most of glucose from digesting sugarm and starch in carbohydrates. Blood sugar
level is controlled by some hormones such as insulin and glucagon. Insulin is a
hormone that control or regulate carbohydrate in the body, lowers the blood sugar
level. Insulin is produce in the beta pancreatic cells. Glucagon is also a hormone that
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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

produced in the pancreas, but have the opposite function from insulin. Glucagon is
used to raise the blood glucose level. Glucose is derived from hexanal, a chain of six
carbon atoms terminating with an aldehyde group. The other five carbon atoms each
bear alcohol groups. Glucose is called an aldohexose. In solution, glucose mainly
exists as the six-membered ring containing a hemiacetal group, which arises from
the reaction of the hydroxy group at C-5 and the aldehyde at C-1. Containing five
carbon atoms and one oxygen atom, this ring is a derivative of pyran. This cyclic
form of glucose is called a glucopyranose, of which two isomers exist.

MATERIAL AND METHOD

Bradford Assay
Materials : Bovine Serum Albumi Standard Set ( 0.125, 0.25, 0.5, 0.75, 1 mg/ml ) vol
20ul, 1X Dye Reagent Vol. 1ml, Cuvettes
20 ul of each standard and unknown sample is pipetted into micocentrifuge
tubes. 1ml of 1X Dye reagent is added to each tube. Using the distilled water and
dye reagent the blank sample is made. The tubes were inverted to mix the solution.
Then it is incubated at room temperature for at least 5 minutes. Samples should not
be incubated longer than 1 hour at room temperature. The spectrophotometer then
was set to 595 nm. The instrument was made to zero with the blank sample.

Ethanol Precipitation
Materials : Laemmli sample buffer, Coomassie blue staining solution ( 50%
methanol, 0.05% (v/v) Coomassie brilliant blur R-250, 10 % (v/v) Acetic Acid, 40%
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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

H2O ), TEMED
125 ul ice cold ethanol and 250 ul serum was added to 1.5 ml microcentrifuge
tube. Then the tube was vortexed and stored in ice bucked for 5 minutes. Next, The
sample was centrifuged at 7000 rpm for 3 minutes at 4 C. The supernatant then was
decated to another mecicentrifuge tube. The sample was labelled Es and stored in
ice bucked. Towel paper was twisted and inserted into the microcentrifuge tube to
draw off the last of the supernatant from above the precipitated proteins bu capilary
action. 1ml chilled 50 % ethanol was added to the precipitate to wash out the last
traces of supernatant. Repeated aspiration is resuspended with 1000 ul automatic
pipette to break up the pellet. This is done quickly so do not heated up preparation
with warmth of your fingers. Then, the sample was centrifuged at 7000 rpm for 3
minutes at 4C. The wash supernatant was decanted into the plastic waste cup. Next,
the last drop was removed with a towel paper. Ep fraction was our precipitate.
Finally, stored in ice bucked.

SDS PAGE sample preparation

Materials : Laemmli sample buffer, Coomassie blue staining solution ( 50%
methanol, 0.05% (v/v) Coomassie brilliant blur R-250, 10 % (v/v) Acetic Acid, 40%
H2O ), TEMED
50 ul ES was taken out to another microcentrifuge tube and 225 ul sample
buffer was added. Then, 450 ul sample buffer was added to our Ep fraction. The
precipitate was vortexed to mix throughly with the sample buffer. 1000 ul automatic
pipette is used for trouble resuspending the pelllet. Both Es and Ep was put into a
rack in the boiling water bath and boiled for 5 minutes. Only the tips of the tube was
contacted with the boiling water. Next, the microcentrifuge tubes was removed from
the boiling water bath. The exterior was dried with a towel paper. The sample was
then centrifuged at room temperature for 3 minutes to pellet any insoluble materials
that may have precipitated during boiling. The sample was ready to be loaded into
the appropriate wells of te gel. 5 12 ul sample was loaded to the gel. The
precipitates was not stired up before loading.

SDS-PAGE
Materials : Laemmli sample buffer, Coomassie blue staining solution ( 50%
methanol, 0.05% (v/v) Coomassie brilliant blur R-250, 10 % (v/v) Acetic Acid, 40%
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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

H2O ), TEMED
A. Preparation of gels
12% separating gel was prepared by adding the following to a 50 ml
breaker ( for 2 gels): 1.28 ml disstilled water, 1.6 ml 30% acrylamide, 1.04
ml 1.5 M Tris- HCL buffer ( pH 8.8 ), 40 ul 10% SDS. A stacking gel was
prepared by adding thw following to a 50 ml breaker ( for 2 gels ) : 0.7 ml
Distilled water, 0.16 30% acrylamide, 0.125 ml 0.5 M Tris HCL buffer
( pH 6.8 ), 15 ul 10% SDS
B. Casting of separating gel
The gel casting was assembled as intructed by the demonstrator, 40 ul 10
% APS solution, 4 ul TEMED was added. Then was swirled gently for
about 5 secconds, 1 ml automatic pipette was used, about 1,8 ml
separating gel solutoin was added carefully to the gel casting mould
avoiding any spillages. The separating gel solution was immediately
overlayed with distilled water. Next, the polymerize was left for about 30
minutes, The distilled water was poured out after confirming with the
demonstrator that the gel has polymerized.
C. Casting of stacking gel
10 ul 10% APS solution and 1 ul TEMED was added, Then was swirled
gently for 5 secconds. 1 ml automatic pipette was used and 0.5 ml
stacking gel was added on top of polymerized separating gel. The comb
was then placed between the glass plates with one end higher than the
other and the comb was carefully pressed down so that the teeth are level
about 5 mm from the top of the separating gel. Next, stacking gel solution
was added if necessary to ensure that the sides of the well are complete.
Left to polymerized about 20 30 minutes. The comb was carefully
removed and the well was washed out at least 3 times with distilled water.

GEL STAINING
Materials : Laemmli sample buffer, Coomassie blue staining solution ( 50%
methanol, 0.05% (v/v) Coomassie brilliant blur R-250, 10 % (v/v) Acetic Acid, 40%
H2O ), TEMED
First the gel was removed and stained with coomasiie blue in wooble table for
1 hour. Then the gel was destained with destain solution in wooble table for 1 our,
the destaining solution was changed every 1 hour with fresh one until background is
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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

clear. Finally, the gel was wrapped in a piece of cellophane plastic.

ELISA
Materials : Rabbit anti human AFP coated microtiter plate with 96 wells; zero
buffter, 13 ml; Reference standard set, contain 0, 5 , 30, 50, 250, 300 ng/ml AFP,
lyophilized; Enzyme Conjugate Reagent, 18 ml; TMB reagent, 11 ml; Stop soultion
(1N HCL0, 11) ml; Distilled water; Microtiter plate reader ( with a bandwidth of 10 nm
or less and an optical density range of 0-2 OD or greater at 450 nm wavelenght ).
First, the desired number of coated wells in the holder was secured. 20 ul of
standard, speciments, and controls was dispensed into appropriate wells. Then 100
ul of zero buffer was dispensed into each well. After that, it was mixed throughly for
30 seconds. Next, was incubated at room temperature for 30 minutes. The
incubation mixture was removed by flicking plate content into waste container. The
microtiter wells was rinsed and flicked 5 times with distilled water. Then the wells was
striked onto absorbent paper or paper towel to remove all residual water droplets.
150 ul of Enzyme Conjugate Reagent was dispensed into each well. Then was
gently mixed for 5 seconds. Next, it was incubated at room temperature for 30
minutes. The incubation mixture was removed by flicking plate content into a waste
container. The microtiter wells was then rinsed and flicked 5 times with distilled
water. The wells was striked sharply onto absorbnt paper to remove residual water
droplets. 100 ul of TMB reagent was dispensed into each well, was mixed for 5
seconds and then was incubated at room temperature for 20 minutes. The reaction
was stopped by adding 100 ul of Stop Solution to each well and was gently mixed for
30 seconds. Finally the optical density was read at 450 nm with a microtiter reader
within 15 minutes.

Determination of blood sugar level

Materials : O-toluidine reagent, standar glucose solution 200mg/dl, distilled water,
centrifuge, spectrophotometer, cuvette, reaction tube, aluminium foil, water bath
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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

Performing standar curve standard glucose solutions
Glucose standar 200 mg/dl are provided. Concentration glucose standar point
was prepared.
Tubes

Concentration

Glucose standar

(mg/dl)
50
80
100
150

(mL)
0.6
0.8
1
1.2

1
2
3
4
Blank
Table 1 Concentration of Glucose Standard

Water (mL)

Total mL per

1.4
1.2
1
0.8
2

tube
2
2
2
2
2

2 mL 1% O-toluidine as added and mixed well. The tube as put in a boiling water
bath for 10 minutes. Finally, the absorbance was measured at 630 nm.
Measuring of blood glucose
Blood is drawn from a vein and transfered into a centrifuge tube. Next, serum
s obtained by centrifugation of blood for 10 minutes. Glucose concentration was
detemined in the provided serum sample using the O-toluidine method as follows :
0.1 ml of distilled water or standard glucose or serum was added in a clean dry test
tube. Then 2 ml of O-toluidine reagent was added.
Blank
Distilled water
0.1 ml
Standard
Test
O-toluidine
1 ml
Table 2 Volume of Standard and Test

Standard
50 ul
1 ml

Test
50 ul
1 ml

The content of each tube was mixed and the tube opening was covered with
aluminium foil. Then the tubes was put in a boiling water bath for 10 minutes. Test
tube was removued from the water bath and cooled under tap water. Next, the
absorbance was read at Imax 630 nm. Finally, the concentration of BSL in the
provided blood samples was calculated using the absorbance reading of standard
glucose.

RESULT
Standard curve of Bradford assay and the result of our sample calculation

Protein

Measurement

Absorbance
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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

concentration (mg/

at 595 nm

mL)
Y
1
2
X
0.125
0.142 0.143
0.1425
0.25
0.295 0.298
0.2965
0.5
0.483 0.488
0.4855
0.75
0.724 0.728
0.726
1
0.815 0.814
0.8145
Table 3 Concentration absorbance of BSA standard test (absorbance at 595
nm)
The absorbance at 595 nm of 5 samples of known concentration is recorder 2 times
each, and the average value is calculated.

Standard Curve for Bradford Test

0.9
f(x) = 0.78x + 0.08
R = 0.98

0.8
0.7
0.6
0.5

Protein Concentration (mg/mL) 0.4

0.3
0.2
0.1
0
0 0.2 0.4 0.6 0.8 1 1.2

Absorbance at 595 nm

Graph 1 Standard Curve

Linear eqquation is
y = 0.779x + 0.083
R2 = 0.976

Sample 1 (A)
Sample 2 (W)

1
0.532
0.673

2
0.539
0.678

Average (x)
0.5355
0.6755

Table 4 Absorbance 595nm of subjects

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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

The absorbance at 595 nm of dilluted serum proteins of both subject is taken twice
and the average value is then calculated
y = 0.779x + 0.083
y (A) = 0.779(0.5355) + 0.083
= 0.4171545 + 0.083
= 0.5001545 x 100 (fp)
= 50.01 mg/ml
y (W) = 0.779(0.6755) + 0.083
= 0.5262145 + 0.083
= 0.6092145 x 100 (fp)
= 60.92 mg/ml

Image of SDS-PAGE gel and Rf of specific band

ES

ES

75
50

Lenght of gel (L)

= 5.6 cm

25

EP

EP

marker

Figure 1 Result of SDS Page gel

SDS Page, showing bands of proteins compared to a marker. Ep and ES fraction
of protein from both subjects are compared to the marker.
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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

MW
S1
75
S2
50
S3
25
S4
z
Table 5 Standard of SDS Page

log
4.875
4.699
4.398
x

d
1.25
1.9
3.4
4.5

L
5.6
5.6
5.6
5.6

Rf (d/L)
0.223
0.339
0.607
0.804

SDS-PAGE Standard Curve

Standard of SDS-PAGE
0.7
0.6

f(x) = - 0.81x + 4.18

R = 0.99

0.5
0.4
Rf 0.3
0.2
0.1
0
4.3

4.4

4.5

4.6

4.7

4.8

4.9

Log

Graph 2 Standard Curve for SDS PAGE

Linear equation is
y = -0.8011x + 4.1232
R2 = 0.994

log = x

= (y 4.1232) 0.8011 x (-1)

= (0.804 4.1232) 0.8011 x (-1)
= -3.3192 0.8011 x (-1)
= -4.143302958 x (-1)
= 4.143302958 cm

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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

MW = z

= antilog x
= antilog 4.143302958
= 13909.2271Da
= 13.9 kDa

Standard curve of ELISA test and the result of your sample calculation
A
0.041
0.086
0.156
0.388
0.800

S1
S2
S3
S4
S5
Table 6 Standard for ELISA

Conc. (ng/ mL)

0.0
5.0
20.0
50.0
150.0

ELISA Standard Curve

Standard Curve of Elisa Test

160
140

f(x) = 196.21x - 12.72

R = 0.98

120
100
Concentration (ng/mL)

80
60
40
20
0
0

0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9

A

Graph 3 Standard curve for ELISA

Linear equation is
y = 196.2x 12.72
A1-1 (1) = 196.2(0.059) - 12.72
= 11.5758 12.72
= - 1.1442 ng/mL
A1-1 (2) = 196.2(0.052) 12.72
= 10.2024 12.72
= - 2.5176 ng/mL
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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

Standard curve of blood sugar determination assay and the result of our
sample calculation
A1
A2
S1
0.194
0.194
S2
0.315
0.318
S3
0.329
0.325
S4
0.428
0.43
Table 7 Standard for blood sugar determination

Mean A
0.194
0.317
0.327
0.429

Conc.
50
80
100
150

BSL assay Standard Curve

Standard Curve of Blood Sugar

160
140
120
100

Conc. (mdg/ dL)

f(x) = 422.9x - 38.95

R = 0.94

80
60
40
20
0
0.15

0.2

0.25

0.3

0.35

0.4

0.45

Graph 4 Standard curve for blood sugar

Linear equation is
y = 423.17x 38.985
R2 = 0.9384
A1
Sampe 1
0.313
Sample 2
0.294
TABLE 8 Blood Sugar Level in Our Groups Samples

A2
0.313
0.300

Mean A :
-

Sample 1 = (0.313+0.313) 2
= 0.313
Sample 2 = (0.294+0.300) 2
= 0.297

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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

Concentration :
-

y = 423.17x 38985
y (sample 1) = 423.17(0.313) 38.985
= 132.45221 38.985
= 93.46721 mg/dL
y (sample 2) = 423.17(0.297) 38.985
= 125.6814 38.985
= 86.69649 mg/dL

DISCUSSION
The Bradford assay uses BSA ( Bovine Serum Albumin ) standard as a
comparison to determine the unknown serum protein concentration. This is because
of the close resemblance of the bovine protein to human blood which mostly consist
of albumin. In this experiment the absorbance reading is done 2 times to avoid the
erorr that can happened. The 2 result dont have a significant difference, this means
that the standard curve is accurate as basis for comparison.
On the calculation of the protein on sample A and B we already know that the
concentration of protein on sample A is 50.01 mg/ml, and the concentration on
sample B is 60.92 mg/ml. Based on the literature, the protein concentration on
sample A is a little bit low compared to the normal plasma concentration that is
between 50 90 mg/ml in a healthy adults, and normal protein concentration on
sample B. The result that happened in sample A can be happened due to errors in
the working method or lack of protein in the blood. The other reason that can cause
this low concentration of protein is the healthiness of the person. Usually, the protein
concentration in the blood can be decreased if the person is not in a healthy
condition or sick. Globulin is the building block for immunoglobulins, the main protein
of the immune system. With immune deficiency, the number of immunoglobulins is
reduced, which decreases blood protein levels.
Malnutrition, overhydration and affect of drugs can also cause low
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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

concentration of protein in blood. If a person's diet lacks protein or any of these
amino acids, his body cannot make albumin and globulin, resulting in low blood
protein. Blood protein levels are measured as the concentration of proteins per
deciliter of blood. With overhydration, the blood volume increases, causing the
protein level to decrease in proportion.

The absolute levels of albumin and globulin

are normal, but the ratio of proteins to fluid has decreased. Several pharmaceuticals
can reduce total blood protein levels.

Among them are estrogens, oral

contraceptives and any drug that is toxic to the liver. Most error is because of human
eror. Improper mixing the dilluted serum protein immediate after injection of
coomassie stain.
In SDS Page, the more intense colouration of the band, the more abundant
that specific protein responsible for that band and the bigger molecular weight of the
protein, the more it will be retained. Ep fraction contains the precipitate proteins from
the result of ethanol precipitation. Es fraction contain protein that do not precipitate
after the ethanol precipitation procedure. These are the proteins found in the
supernatant.
Ep band travels further than the Es band determine by the distance they
travelled from loading the well. The small molecules of protein will migrate faster
than the larger molecules. SDS interaction with the protein cause the protein
migrates to the anode because of the overall negative charge.
As we can see in the result of SDS PAGE gel, there was a thick band around
50 molecular weight. The thick band can be interpreted that albumin is the major
protein blood plasma so it can be conjectured that the dark and heavy band in both
Ep and Es bands of both subjects are caused by the abundant albumin. The thick
bands means that there is same protein in both subjects. There is also possible that
the band is not in a straight line. This is happened because of error in preparing the
gel and the polymerization is not event in the gel. The different polymerization make
the result that same bands are bent line.
The aim of ELISA test is to determine of the Antigen AFP concentration in
human serum. ELISA test are utilized to detect substances that have antigenic
properties, primarily proteins. The concentration of AFP of sample A is -1.1442 ng/mL
and for sample B is -2.5176. Compared to the normal range of AFP concentration
that is <20 ng/mL the cocentration of AFP in sample A and B is normal. In some
cases the concentration of AFP can be elevated when hepatocellularcarcinoma or
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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

other type of tumour happened.
The blood sugar level test is used to measure blood glucose from serum
sample. In this method O-toluidine is used that react in hot glacial acetic acid with
the terminalaldehyde group of glucose to produce a blue green colored condensation
product that can be measured colorimetrically at Imax 630 nm. Our result of Sample
A blood sugar level is 93.46721 mg/dl and 86.69649 mg/dL for sample B. According
to the normal range of blood glucose level that is between 82 to 110 mg/dL in normal
fasting, our result is in normal range. To increase the accuracy of the test, it is
necessary for the students in a fasting period. The blood sugar level can be elevated
in some diseases such as hyperthyroidism, pancreatic cancer, diabetes, etc and can
also be low in some cases such as drinking alcohol, side effect of drugs, etc.
Conclution for this experiment is that the instrument is highly necessary for
doing the experiment. Such as spectrophotometer, pipette, etc. Most of the erorrs
that happened in the experimet is based on the human erorrs. High concentration
and accuracy are required to get a successful result at the end.

REFERENCE

Definition of protein, Available at :

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Proteomic Report A-1-1 Adrian Jayadi (07120100005)

http://www.medterms.com/script/main/art.asp?articlekey=6554

Stucture of Protein, Available at:

http://themedicalbiochemistrypage.org/protein-structure.html#primary
Bradford Protein Concentration Assay, Available at :
http://ww2.chemistry.gatech.edu/~lw26/bCourse_Information/4581/techniques/bradfo
rd/bradford.html

SDS PAGE, Available at :

http://askabiologist.asu.edu/sds-page
What are the Cause of Low Blood Protein?, Available at :
http://www.ehow.com/about_5132551_causes-low-blood-protein.html
Blood Sugar, Available at :
http://en.wikipedia.org/wiki/Blood_sugar
Ethanol Precipitation of Protein, Available at :

http://www.kendricklabs.com/EtOHppt_2008.pdf
Glucose, Available at :
http://en.wikipedia.org/wiki/Glucose
Anonymous. Laboratory protocol for Fundamental Medical Science 1. Faculty

of Medicine Universitas Pelita Harapan : Tangerang;2010

Murray, Robert K. Harpers Illustrated Biochemistry. 27 th ed. Singapore:

McGraw- Hill; 2006.

Sherwood Lauralee 2007, Human Physiology From cells to Systems, 6 th

edition, Thomson Brooks/Cole, USA

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