1

HUMAN CLUSTERIN ELISA

4.

Cat. No.: BA3001

In the Human Clusterin ELISA, standards, quality controls and samples are
incubated in microplate wells pre-coated with monoclonal anti-humanclusterin antibody. After 60 minutes incubation and washing, biotin labelled
second monoclonal anti-human clusterin antibody is added and incubatedwith captured clusterin for 60 minutes. After another washing,streptavidin-HRP conjugate is added. After 30 minutes incubation and thelast washing step, the remaining conjugate is allowed to react with thesubstrate solution (TMB). The reaction is stopped by addition of acidic
solution and absorbance of the resulting yellow product is measured.
The absorbance is proportional to the concentration of clusterin. A standard
curve is constructed by plotting absorbance values against concentrationsof standards, and concentrations of unknown samples are determined using
this standard curve.

For research use only!
1.

INTENDED USE

The BA1029 Human Clusterin ELISA is a sandwich enzyme immunoassay forthe quantitative measurement of human clusterin.
FeaturesIt is intended for research use only
The total assay time is less than 3.5 hours
The kit measures clusterin in serum, plasma (EDTA, citrate,
heparin), cerebrospinal fluid (CSF) and urine
Assay format is 96 wells
Quality Controls are human serum basedStandard is human serum based native proteinComponents of the kit are provided ready to use, concentrated orlyophilized

5.

PRECAUTIONS

For professional use only
Wear gloves and laboratory coats when handlingimmunodiagnostic materials
Do not drink, eat or smoke in the areas where immunodiagnostic
materials are being handledThis kit contains components of human origin. These materials
were found non-reactive for HBsAg, HCV antibody and for HIV 1/2
antigen and antibody. However, these materials should be handled
as potentially infectious, as no test can guarantee the complete
absence of infectious agents
Avoid contact with the acidic Stop Solution and Substrate Solution,
which contains hydrogen peroxide and tetramethylbenzidine(TMB). Wear gloves and eye and clothing protection when handlingthese reagents. Stop and/or Substrate Solutions may cause
skin/eyes irritation. In case of contact with the Stop Solution and
the Substrate Solution wash skin/eyes thoroughly with water andseek medical attention, when necessary
The materials must not be pipetted by mouth

6.

TECHNICAL HINTS






Reagents with different lot numbers should not be mixed
Use thoroughly clean glassware
Use deionized (distilled) water, stored in clean containers
Avoid any contamination among samples and reagents. For thispurpose, disposable tips should be used for each sample andreagent
Substrate Solution should remain colourless until added to theplate. Keep Substrate Solution protected from light
Stop Solution should remain colourless until added to the plate.
The colour developed in the wells will turn from blue to yellow
immediately after the addition of the Stop Solution. Wells that are
green in colour indicate that the Stop Solution has not mixed
thoroughly with the Substrate Solution
Dispose of consumable materials and unused contents inaccordance with applicable national regulatory requirements


2.

STORAGE, EXPIRATIONStore the complete kit at 2-8°C. Under these conditions, the kit is stable until
the expiration date (see label on the box).
For stability of opened reagents see Chapter 9.
3.

Areas of investigation:
Coronary heart diseases
Myocardial infarction
Neurodegenerative diseases
Kidney degenerative disease
Renal tubular damage
v_01

INTRODUCTION

Clusterin (Apolippoprotein J; SP-40,40; TRPM-2; SGP-2; pADHC-9; CLJ; T64;
GP III; XIP8) is a highly conserved disulfide-linked secreted heterodimericglycoprotein of 75-80 kDa but truncated forms targeted to nucleus havealso been identified.
The protein is constitutively secreted by a number of cell types including
epithelial and neuronal cells and is a major protein in physiological fluids
including plasma, milk, urine, cerebrospinal fluid and semen. Due to its wide
tissue distribution many diverse physiological functions have been
attributed to clusterin including sperm maturation, membrane recycling,
lipid transportation, tissue remodelling, complement inhibition and cell-cellor cell-substratum interactions. Moreover, it was propose, that clusterin
functions is as an extra cellular chaperon that stabilizes stressed proteins ina folding-competent state and protein has also been implicated inprogrammed cell death. Another defining prominent of clusterin is its
induction in many severe physiological disturbances states including kidneydegenerative diseases, prostate and vesicle carcinogenesis, ovarian cancer
and several neurodegenerative conditions (Alzheimer’s disease).
Recent study demonstrate, that serum clusterin level increases significantly
in diabetic type II patients and in patients with developing coronary heartdisease, or myocardial infarction. These date raise the possibility thatelevated clusterin levels in serum may represent a strong indication ofvascular damage.
In patients with systemic lupus erythematosus (SLE) was found reduced
serum clusterin levels that correlated inversely with disease activity.
Lowered clusterin levels could be involved in the pathogeneses of SLE on
account of decreased protective effects.
Another interesting observations obtain in rat model suggest thatmeasurement of urinary clusterin levels may be a useful clinical valuable
marker for the severity of renal tubular damage. Furthermore, urinary
clusterin may also help to differentiate between tubular and glomerular
forms of proteinuria.

TEST PRINCIPLE






2

7.

REAGENT SUPPLIED
State

Quantity

Antibody Coated Microtiter Strips

ready to use

96 wells

Biotin Labelled Antibody Conc. (50x)concentrated

0.26 ml

Streptavidin-HRP Conjugate

ready to use

13 ml

Master Standard

lyophilized

2 vials

Quality Control HIGH

lyophilized

2 vials

Quality Control LOW

lyophilized

2 vials

Biotin-Ab Diluent

ready to use

13 ml

Dilution Buffer

ready to use

50 ml

Wash Solution Conc. (10x)concentrated

100 mlSubstrate Solution

ready to use

13 ml

Stop Solution

ready to use

13 ml

Kit Components

8.

MATERIAL REQUIRED BUT NOT SUPPLIED

Deionized (distilled) water
Test tubes for diluting samples
Glassware (graduated cylinder and bottle) for Wash Solution(Dilution Buffer)
Precision pipettes to deliver 5-1000 l with disposable tips
Multichannel pipette to deliver 100 l with disposable tips
Absorbent material (e.g. paper towels) for blotting the microtitrate
plate after washing
Vortex mixer
Orbital microplate shaker capable of approximately 300 rpm
Microplate washer (optional). [Manual washing is possible but notpreferable.]
Microplate reader with 450  10 nm filter, preferably withreference wavelength 630 nm (alternatively another one from theinterval 550-650 nm)
Software package facilitating data generation and analysis(optional)










9.

PREPARATION OF REAGENTS

All reagents need to be brought to room temperature prior to use.
Centrifuge liquid containing microtube vials before opening.
Always prepare only the appropriate quantity of reagents for your test.
Do not use components after the expiration date marked on their label

Streptavidin-HRP Conjugate
Biotin-Ab Diluent
Dilution Buffer
Substrate Solution
Stop Solution
Stability and storage:
Opened reagents are stable 3 month when stored at 2-8°C.

Assay reagents supplied concentrated or lyophilized:
Human Clusterin Master Standard
Refer to the Certificate of Analysis for current volume of Dilution Buffer
needed for reconstitution of standard!!!
Reconstitute the lyophilized Master Standard with Dilution Buffer just prior
to the assay. Let it dissolve at least 15 minutes with occasional gentle
shaking (not to foam). The resulting concentration of the clusterin in thestock solution is 160 ng/ml.
Prepare set of standards using Dilution Buffer as follows:
Volume of Standard

Dilution Buffer

Concentration

Stock

-

160 ng/ml

250 µl of stock

250 µl80 ng/ml

250 µl of 80 ng/ml

250 µl40 ng/ml

250 µl of 40 ng/ml

250 µl20 ng/ml

250 µl of 20 ng/ml

250 µl10 ng/ml

250 µl of 10 ng/ml

250 µl5 ng/ml

Prepared Standards are ready to use, do not dilute them.
Stability and storage:
Do not store the Standard stock solution and set of standards.
Quality Controls HIGH, LOW
Refer to the Certificate of Analysis for current volume of Dilution Buffer
needed for reconstitution and for current Quality Control concentration!!!
Reconstitute each Quality Control (HIGH and LOW) with Dilution Buffer justprior to the assay. Let it dissolve at least 15 minutes with occasional gentle
shaking (not to foam).
Reconstituted Quality Controls are ready to use, do not dilute them.
Stability and storage:
Do not store the reconstituted Quality Controls.
Note:
Concentration of analyte in Quality Controls need not be anyhow associatedwith normal and/or pathological concentrations in serum or another body
fluid. Quality Controls serve just for control that the kit works in accordancewith IFU and CoA and that ELISA test was carried out properly.

Assay reagents supplied ready to use:

Biotin Labelled Antibody Conc. (50x)Antibody Coated Microtiter Strips

Prepare the working Biotin Labelled Antibody solution by adding 1 part
Biotin Labelled Antibody Concentrate (50x) with 49 parts Biotin-Ab Diluent.
Example: 20 µl of Biotin Labelled Antibody Concentrate (50x) + 980 µl of
Biotin-Ab Diluent for 1 strip (8 wells).

Stability and storage:
Return the unused strips to the provided aluminium zip-sealed bag withdesicant and seal carefully. Remaining Microtiter Strips are stable 3 monthstored at 2-8°C and protected from the moisture.

v_01

Stability and storage:
Opened Biotin Labelled Antibody Concentrate (50x) is stable 3 months whenstored at 2-8°C.
Do not store the diluted Biotin Labelled Antibody solution.

3

Wash Solution Conc. (10x)9.

Dilute Wash Solution Concentrate (10x) ten-fold in distilled water toprepare a 1x working solution. Example: 100 ml of Wash Solution
Concentrate (10x) + 900 ml of distilled water for use of all 96-wells.

10.

Stability and storage:
The diluted Wash Solution is stable 1 month when stored at 2-8°C. Opened
Wash Solution Concentrate (10x) is stable 3 months when stored at 2-8°C.
10.

Wash the wells 5-times with Wash Solution (0.35 ml per well). After
final wash, invert and tap the plate strongly against paper towel.
Add 100 µl of Substrate Solution into each well. Avoid exposing the
microtiter plate to direct sunlight. Covering the plate with e.g.
aluminium foil is recommended.
Incubate the plate for 10 minutes at room temperature. The
incubation time may be extended [up to 20 minutes] if the reaction
temperature is below than 20°C. Do not shake the plate during the
incubation.
Stop the colour development by adding 100 µl of Stop Solution.
Determine the absorbance of each well using a microplate reader
set to 450 nm, preferably with the reference wavelength set to
630 nm (acceptable range: 550-650 nm). Subtract readings at
630 nm (550-650 nm) from the readings at 450 nm. The
absorbance should be read within 5 minutes following step 12.

11.

12.
13.

PREPARATION OF SAMPLESThe kit measures human clusterin in serum, plasma (EDTA, citrate, heparin),
CSF and urine.
Samples should be assayed immediately after collection or should be stored
at -20°C. Mix thoroughly thawed samples just prior to the assay and avoid
repeated freeze/thaw cycles, which may cause erroneous results. Avoid
using hemolyzed or lipemic samples.
Serum or plasma samples:
Dilute samples just prior to perform the test 3 000x with Dilution Buffer in
two steps as follows:
Dilution A (50x):
Add 5 µl of sample into 245 µl of Dilution Buffer and mix well (not to foam).
Vortex is recommended.
Dilution B (60x):
Add 5 µl of Dilution A into 295 µl of Dilution Buffer to prepare final dilution
3 000x. Mix well (not to foam). Vortex is recommended.

Note 1: If some samples and standard/s have absorbances above the upper
limit of your microplate reader, perform a second reading at 405 nm. A new
standard curve, constructed using the values measured at 405 nm, is used
to determine clusterin concentration of off-scale standards and samples.
The readings at 405 nm should not replace the readings for samples thatwere “in range” at 450 nm.
Note 2: Manual washing: Aspirate wells and pipet 0.35 ml Wash Solution
into each well. Aspirate wells and repeat four times. After final wash, invert
and tap the plate strongly against paper towel. Make certain that Wash
Solution has been removed entirely.

strip 1+2

strip 3+4

strip 5+6

strip 7+8

strip 9+10 strip 11+12

Blank

CSF samples:A

Sample 8

Dilute CSF samples just prior to perform the test 100x with Dilution Buffer,e.g. 5 µl of sample + 495 µl Dilution Buffer. Mix well (not to foam). Vortex
is recommended.

Standard
160

Sample16

Sample24

Sample32

B

Standard
Sample 1 Sample 980

Sample17

Sample25

Sample33

CStandard
Sample 2
40

Sample10

Sample18

Sample26

Sample34

D

Standard
Sample 320

Sample11

Sample19

Sample27

Sample35

EStandard
Sample 4
10

Sample12

Sample20

Sample28

Sample36

FStandard 5 Sample 5

Sample13

Sample21

Sample29

Sample37

G

QC HIGH

Sample 6

Sample14

Sample22

Sample30

Sample38

H

QC LOW

Sample 7

Sample15

Sample23

Sample31

Sample39

Urine samples:
Dilute urine samples just prior to perform the test 10x with Dilution Buffer,e.g. 15 µl of sample + 135 µl of Dilution Buffer for singlets or 25 µl of sample+ 225 µl of Dilution Buffer for duplicates. Mix well (not to foam). Vortex isrecommended.
Stability and storage:
Serum and plasma samples should be stored at -20°C, or preferably at -70°C
for long-term storage. CSF and urine samples should be stored at -70°C.
Do not store the diluted samples.
See Chapter 13 for stability of serum and plasma samples when stored at 28°C, effect of freezing/thawing and effect of sample matrix (serum/plasma)on the concentration of human clusterin.
Note: It is recommended to use a precision pipette and a careful techniqueto perform the dilution in order to get precise results.
11.

ASSAY PROCEDURE

1.

Pipet 100 µl of diluted Standards, Quality Controls, Dilution Buffer(=Blank) and samples, preferably in duplicates, into the appropriate
wells. See Figure 1 for example of work sheet.
Incubate the plate at room temperature (ca. 25°C) for 1 hour,
shaking at ca. 300 rpm on an orbital microplate shaker.
Wash the wells 5-times with Wash Solution (0.35 ml per well). After
final wash, invert and tap the plate strongly against paper towel.
Add 100 µl of Biotin Labelled Antibody solution into each well.
Incubate the plate at room temperature (ca. 25°C) for 1 hour,
shaking at ca. 300 rpm on an orbital microplate shaker.
Wash the wells 5-times with Wash Solution (0.35 ml per well). After
final wash, invert and tap the plate strongly against paper towel.
Add 100 µl of Streptavidin-HRP Conjugate into each well.
Incubate the plate at room temperature (ca. 25°C) for 30 min,shaking at ca. 300 rpm on an orbital microplate shaker.

2.
3.
4.
5.
6.
7.
8.
v_01

Figure 1: Example of a work sheet.
12.

CALCULATIONS

Most microplate readers perform automatic calculations of analyte
concentration. The standard curve is constructed by plotting the mean
absorbance (Y) of Standards against the known concentration (X) of
Standards in logarithmic scale, using the four-parameter algorithm. Results
are reported as concentration of clusterin ng/ml in samples.
Alternatively, the logit log function can be used to linearize the standard
curve, i.e. logit of the mean absorbance (Y) is plotted against log of the
known concentration (X) of Standards.
The measured concentration of samples calculated from the standard
curve must be multiplied by their respective dilution factor, because
samples have been diluted prior to the assay, e.g. 20 ng/ml (from standard
curve) x 3 000 (dilution factor) = 60 000 ng/ml = 60 µg/ml.

4

Human Clusterin ELISA
Standard Curve

Absorbance (A450 nm-A630nm)

3.5
3.0
2.5
2.0
1.5

Mammalian
serum sample

Observedcrossreactivity

Mammalian
serum sample

Observedcrossreactivity

Bovineno

Monkey

yes

Cat

no

Mouse

no

Dog

no

Pig

no

Goat

no

Rabbitno

Hamster

no

Rat

no

Horse

no

Sheep

no

Presented results are multiplied by respective dilution factor.

1.0

Precision
0.5

Intra-assay (Within-Run) (n=8)

0.0
1

10

100

SampleMean (µg/ml)

SD (µg/ml)

CV (%)

1

68.0

4.8

7.0

2

103.6

5.6

5.4

1000

Concentration of Hu Clusterin (ng/ml)

Figure 2: Typical Standard Curve for Human Clusterin ELISA.

13.

Inter-assay (Run-to-Run) (n=8)
SampleMean (µg/ml)

SD (µg/ml)

CV (%)

1

62.4

5.3

8.5

2

95.8

6.7

7.0

PERFORMANCE CHARACTERISTICS

Typical analytical data of Human Clusterin ELISA are presented in thischapter.
Spiking Recovery
Sensitivity
Limit of Detection (LOD) (defined as concentration of analyte giving
absorbance higher than mean absorbance of blank* plus three standard
deviations of the absorbance of blank: Ablank + 3xSDblank) is calculated fromthe real human clusterin values in wells and is 0.5 ng/ml.
*Dilution Buffer is pipetted into blank wells.
Limit of assay
Results exceeding clusterin level of 160 ng/ml should be repeated with more
diluted samples. Dilution factor needs to be taken into consideration in
calculating the clusterin concentration.
Results from assaying of urine samples exceeding clusterin level of 40 ng/ml
should be repeated with more diluted urine samples for correct values.
Dilution factor needs to be taken into consideration in calculating theclusterin concentration.
Specificity
The antibodies used in this ELISA are specific for human clusterin.
Sera of several mammalian species were measured in the assay. See results
below.
For details, please contact us at info@biolabassays.eu.

v_01

Serum samples were spiked with different amounts of clusterin andassayed.
SampleObserved(µg/ml)Expected(µg/ml)Recovery O/E
(%)

1

11.0
50.9
31.6
21.5

51.0
31.0
21.0

99.8
102.1
102.5

2

21.5
61.8
43.6
34.4

61.5
41.5
31.5

100.4
105.2
109.2

Linearity
Serum samples were serially diluted with Dilution Buffer and assayed.

SampleDilution

1

2x
4x
8x

2

2x
4x
8x

Observed(µg/ml)
49.2
24.2
12.8
6.1
70.3
33.5
17.3
8.9

Expected(µg/ml)
24.6
12.3
6.2
35.1
17.6
8.8

RecoveryO/E (%)
98.4
103.8
98.7
95.4
98.2
101.2

5

Effect of sample matrix
Heparin, citrate and EDTA plasmas were compared to respective serum
samples from the same 10 individuals. Results are shown below:
Volunteer
No.

Serum
(µg/ml)Sample1

Plasma (µg/ml)

EDTA

CitrateHeparin

-20°C

99.6

199.4

72.9

89.6

2-8°C, 1 day

68.2

189.9

63.8

104.0

2-8°C, 7 days

72.6

209.7

65.1

158.1

-20°C

71.6

188.0

48.6

58.9

2-8°C, 1 day

101.7

178.8

63.0

81.3

235.5

84.6

99.0

1

140.4

2

127.7

179.2

113.0

104.9

3

126.8

170.5

106.0

107.8

2-8°C, 7 days

102.3

4

113.4

157.6

96.3

92.5

-20°C

71.9

155.6

54.0

72.5

5

130.8

166.2

108.5

113.1

2-8°C, 1 day

62.4

165.5

54.6

87.1

6

85.5

109.8

57.3

74.3

2-8°C, 7 days

75.1

137.0

58.7

97.4

7

82.4

126.7

73.7

86.3

8

85.3

114.8

64.1

74.0

9

88.0

109.3

59.2

82.7

10

89.4

149.1

79.1

74.4

Mean (µg/ml)

107.0

146.1

86.6

94.0

-

136.6

81.0

87.8

-

0.83

0.89

0.89

determination R2

Heparin
129.7

Serum
(µg/ml)

EDTA
177.5

Mean
Plasma/Serum(%) ofCoefficient

Citrate108.9

Plasma (µg/ml)

Incubation
Temp,
Period2

3Effect of Freezing/Thawing
No decline was observed in concentration of human clusterin in serum andplasma samples after repeated (5x) freeze/thaw cycles. However it isrecommended to avoid unnecessary repeated freezing/thawing of thesamples.
Sampl
e

Number off/t cyclesSerum(µg/ml)1x
1Effect of Sample Matrix
250

C oncentration of H u C lusterin (µ g/m l)

Serum
EDTA Plasma

200

Citrate Plasma

2

Heparin Plasma
150

100

3
50

0

14.
1

2

3

4

5

6

7

8

9

Plasma (µg/ml)
EDTA

CitrateHeparin

104.7

145.7

58.1

89.0

3x

91.1

166.2

58.3

73.2

5x

96.3

186.2

64.2

89.7

1x

78.2

145.7

51.2

56.6

3x

95.2

149.5

57.3

87.4

5x

84.1

186.6

56.1

72.9

1x

102.5

168.9

60.9

85.7

3x

99.3

209.0

71.3

80.3

5x

108.8

208.9

56.0

113.6

DEFINITION OF THE STANDARD

10Volunteers
Figure 3: Clusterin levels measured using Human Clusterin ELISA from10
individuals using serum, heparin, citrate and EDTA plasma, respectively.
Stability of samples stored at 2-8°C
Samples should be stored at –20°C. However, no decline in concentration ofclusterin was observed in serum and plasma samples after 7 days whenstored at 2-8°C. To avoid microbial contamination, samples were treatedwith -aminocaproic acid and sodium azide, resulting in the final
concentration of 0.03% and 0.1%, respectively.

Standard in this assay is human serum based native clusterin. Native
clusterin is 75-80 kDa heterodimeric glycoprotein. 1ng/ml of human serum
based standard corresponds to 1ng/ml of recombinant human clusterin
which was used in Biovendor Clusterin ELISA.
15.

PRELIMINARY POPULATION AND CLINICAL DATA

The following results were obtained when serum samples from 200
unselected donors (100 female + 100 male) 4-80 years old were assayedwith the Human Clusterin ELISA in our laboratory.
Normal value
Population Mean (mean +/- SEM): 95.0 +/- 1.5 µg/ml
Normal Range (mean +/- 2SD): 95.0 +/- 42.2 µg/ml
Reference range
The data quoted in these instructions should be used for guidance only. It is
recommended that each laboratory include its own panel of control sample
in the assay. Each laboratory should establish its own normal andpathological references ranges for clusterin levels with the assay.

v_01

6

High signal and background in all wells
Age and sex dependent distribution of clusterin

Sex

Men

Women

Mean

SD

Min.

Max.

Age
years

n

4-19

17

89.9

16.4

62.5

106.5

20-29

20

88.6

19.8

52.5

132.5

30-39

15

96.6

21.6

56.0

147.0

40-49

91.9

50-59

15
15

88.1

20.9
15.4

67.5
64.0

131.5
107.0

60-80

18

97.8

20.7

71.5

135.0

3-19

17

93.0

23.0

64.0

157.5

20-29

20

97.2

14.8

51.5

113.0

30-39

15

99.6

17.5

64.5

133.5

40-49

15

108.1

22.4

80.5

164.0

50-59

16
17

95.2
94.9

25.9
35.0

59.0
54.0

127.5
164.5

60-80

Clusterin (µg/ml)

Possible explanations:
Improper or inadequate washing
Overdeveloping; incubation time with Substrate Solution should bedecreased before addition of Stop SolutionIncubation temperature over 30°C
High coefficient of variation (CV)
Possible explanation:Improper or inadequate washing
Improper mixing Standards, Quality Controls or samples
18.

References to clusterin:Age and Sex Dependent Distribution of Hu ClusterinC oncentration of H u C luste rin (µ g/m l)

200
180

men

160

women



140



120
100
80



60
40
20



0
0

10

20

30

40

50

60

70

80

90



Age (years)
Figure 4: Clusterin concentration plotted against donor age.

16.

METHOD COMPARISON

BioLab Assays Human Clusterin ELISA has not been compared to any otherimmunoassay.
17.

TROUBLESHOOTING AND FAQS

Weak signal in all wells
Possible explanations:Omission of a reagent or a stepImproper preparation or storage of a reagent

Assay performed before reagents were allowed to come to room
temperatureImproper wavelength when reading absorbance
v_01

REFERENCES

Choi-Miura NH, Oda T: Relationship between multifunctional protein
Clusterin and Alzheimer disease. Neurobiol. Aging 1996; 17(5): 717722
Newkirk MM, Apostolakos P, Neville C and Fortin PR: Systemic lupus
erythematosus, a disease associated with low levels of Clusterin/ApoJ,
and anti-inflammatory protein. J. Rheumatol.1999; 3:597-603
Morrissey C, Lakins J, Moquin A, Hussain M, Tenniswood M:
An antigen capture assay for the measurement of serum Clusterin
concentrations. J. Biochen. Biophys. Methods 2001; 48:13-21
Girton RA, Sundin DP, and Rosenberg ME: Clusterin protects renal
tubular epithelial cells from gentamicin-mediated cytotoxicity. Am J
Renal Physiol. 2002; 282:F709-F709
Trougakos IP, Poulakou M, Stathatos M, Chalikia A, Melidonis A, Gonos
ES:
Serum
levelsofthesenescencebiomarker
Clusterin/apolipoprotein J increase significantly in diabetes type II and
during development of coronary heart disease or at myocardial
infarction. Ex. Gerontology 2002; 37: 1175-1187
Jones SE, Jomary C: Molecules in focus Clusterin. The International J.
of Bioch.  Cell Biol. 2002; 34:427-431
Hidaka S, Kränzlin B, Gretz N, Witzgall R: Urinary Clusterin levels in the
rat correlate with the severity of tubular damage and may help todifferentiate between glomerular and tubular injuries. Cell Tissue Res.
2002; 310:289-296
Ghiggeri GM, Bruschi M, Candiano G, Rastaldi MP, Scolari F, Passerini
P, Musante L, Pertica N, Caridi G, Ferrario F, Perfumo F, and Ponticelli
C: Depletion of clusterin in renal diseases causing nephrotic syndrome.
Kidney Intern. 2002; 62:2184-2194
Chen X, Halberg RB, Ehrhardt WM, Torrealba J and Dove WF: Clusterin
as a biomarker in murine and human intestinal neoplasia. PNAS 2003;
100:9530-9535
Zhang LY, Ying WT, Mao YS, He HZ, Liu Y, Wang HX, Liu F, Wang K,
Zhang DC, Wang Y, Wu M, Qian XH and Zhao XH: Loss of Clusterin bothin serum and tissue correlates with the tumorogenesis of esophageal
squamous cell carcinoma via proteomics approaches. World J
Gastroenterol 2003; 9:650-654
Wang L, Erling P, Bengtsson AA, Truedsson L, Sturfelt G, Erlinge D:
Transcriptional down-regulation of the platelet ADP receptory P2Y12
and Clusterin in patients with systemic lupus erythematosus. J. of
Thromb. And Haemost. 2004; 2:1436-1442
Patel NV, Wei M, Wong A, Finch CE, Morgan TE: Progressive changes
in regulation of apolipoproteins E and J in glial cultures during
postnatal development and aging. Neuroscience Letters 2004;
371:199-204
Krijnen PAJ, Cillessen SAGM, Manoe R, Muller A, Visser CA, Meijer
CJLM, Musters RJP, Hack CE, Aarden LA, and Niessen HWM: Clusterin:a protective mediator for ischemic cardiomyocytes? Am J Physiol
Heart Circ Physiol 2005; 289:H2193-H2202
Kim BM, Kim SY, Lee S, Shin YJ, Min BH, Bendayan M, Park IS: Clusterin
induces differentiation of pancreatic duct calls into insulin-secreting
cells. Diabetologia 2006; 49:311-320
Kruger S, Mahnken A, Kausch I, Feller AC: Value of Clusterin
immunoreactivity as a predictive factor in muscle-invasive urothelial
bladder carcinoma. Urology 2006; 67:105-109

7







Rodriguez-Pineiro AM, De la Cadena MP, Lopez-Saco A, and RodriguezBerrocal FJ: Differential Expression of serum clusterin isoforms incolorectal cancer. Mol. And Cel. Proteomics 2006; 5:1647-1657
Strochi P, Smith MA, Perry G, Tamagno E, Danni O, Pession A, Gaiba A,
Dozza B: Clusterin up-regulation following sub-lethal oxidative stressand lipid peroxidation in human neuroblastoma cells. Neurobiol. of
Aging 2006; 27:1588-1594
Ishii A, Sakai Y, and Nakamura A: Molecular pathological evaluation ofclusterin in a rat model of unilateral ureteral obstruction as a possible
biomarker of nephrotoxicity. Toxicologic Pathology 2007; 35:376-382
Stoop MP, Dekker LJ, Titulaer MK, Burgers PC, Sillevis Smitt PAE, Luider
TM, and Hintzen RQ: Multiple sclerosis-related identified in
cerebrospinal fluid by advanced mass spectrometry. Proteomics 2008;
8:0000-0000

19.

EXPLANATION OF SYMBOLS

Catalogue number
Batch code
CautionUse by dateTemperature limit
References to this product
Manufacturer














Stejskal D, Fiala R: Evaluation of serum and urine Clusterin as apotential tumor marker for urinary bladder cancer. Neoplasma 2006;
53:343-346
Zhang F, Sha J, Wood TG, Galindo CL, Garner HR, Burkart MF, Suarez
G, Sierra JC, Agar SL, Peterson JW, Chopra AK: Alteration in theactivation state of new inflammation-associated targets byphospholipase A.2-activating protein (PLAA). Cellular Signalling 2008;
20:844-861
Zhang F, Suarez G, Sha J, Sierra JC, Peterson JW, Chopra AK:
Phospholipase A.2-activating protein (PLAA) enhances cisplatininduced apoptosis in HeLa cells. Cellular Signalling 2009; 21:1085-1099
Vareka I, Stejskal D, Varekova R, Burianova K, Hnatek J,: Changes inclusterin serum concentration levels in oncologic patients during thecourse of Spa therapy – a pilot study. Biomed Pap Med Fac Univ
Palacky Olomouc Czech Repub. 2009; 153:117-120
Miyake H, Muramaki M, Furukawa J, Kurahashi T, and Fujisawa M:
Serum level of clusterin and its density in men with prostate cancer asnovel biomarkers reflecting disease extension. J Urology 2009; 75:454459
Aguilar-Mahecha A, Cantin C, O'Connor-McCourt M, Nantel A, andBasik M: Development of reverse phase protein microarrays for thevalidation of clusterin, a mid-abundant blood biomarker. Proteome
Science 2009; 7:15
Wang Y, Liu YH, Mai SJ, He LJ, Liao YJ, Deng HX, Guan XY, Zeng YX, Kung
HF and Xie D: Evaluation of serum clusterin as a surveillance tool forhuman hepatocellular carcinoma with hepatitis B virus related
cirrhosis. J Gastr. and Hep. 2010; 25:1123-1128
Thambisetty M, and coll.: Association of plasma clusterin
concentration with severity, pathology, and progression in Alzheimer
Disease. Arch Gen Psychiatry 2010; 67:739-748

For more references on this product see our web pages atwww.biolabassays.eu.

v_01

Read electronic instructions foruse – eIFU
The content is sufficient for 96
tests
Biological risks

20.

ASSAY PROCEDURE - SUMMARY