NEPHSTAR

Complementa (C3) Kit

Catalog No.
1.

DK033

Intended Use

This product is used on NEPHSTAR protein analysis system

for quantitative determination of human Complement 3 ( C 3 ) in
seriin-! as an aid in c'iagnosis of abnorma! C 3 metabolism.

possible to prevent haemolysis. S e r a may be stored at 2-8C

for 48 hours, othenwise freeze at - 2 0 C or below/; do not freeze
and thaw sera more than once. Sample dilutions should be
freshiy prepared on the day of assay. Testing of the following
types of sera may result in misleading vales:
7.1 Highiy lipemic, turbid and haemolysed samples are not
suitable for nephelometric a s s a y s and should not be used
unless centrifuged or prepared using other methods. If the
background is too turbid and can not be removed, please
think of other measuring method.
7.2 Testing of samples containing rheumatoid factors,
paraprctens or circulating immunocomplexes can result in
misleading vales due to non-specific scattering light
possibly generated by these articles.
8.

2. S u m m a r y
C 3 is the central point of the classic and alternative
complement pathway. On activation spilt products of C 3 h a s
important biological functions. C 3 is an opsonin and involved in
immune adherence. C 3 is an anaphyiatoxin and a chemotoxin.
C 3 behaves aiso like an acute phase protein, therefore
increased levis may be found in acute inflammatory reactions.
Decreased levis are reported in complex disease, recurrent
immune
infections
with
pyrogenic
bacteria,
various
glomerulonephritides and in congenital deficiencies.

3. T e s t Principie
Immunonephelometry
is applied. This method nvolves
measuring the light scattered by insoluble complexes formed
by reaction betv^/een specific protein in samples and its
respective antiserum, and the amount of scattered light is
directly proportional to the concentration ot the protein under
condition that antiserum is In e x c e s s . Concentrations are
automaticaliy calculated by reference to a calibration curve
sfored in the instrument.
4.

DB033
DC033
DM033

5.
5.1
5.2
5.3
5.4
5.5
5.6

ame
C 3 Antiserum
C 3 Reaction buffer
C 3 Magnetic card
C 3 Control
Manual

8.1
8.2

8.3

8.4

8.5

Volume/Quantity
1 x 2 . 0 mi
1

1x0.3mL

8.7

Materials required but not supplied

N E P H S T A R Protein analysis system ( N S 1 0 0 )
N E P H S T A R Accessory pack ( D K 1 1 0 )
Electronic pipette ( Y B 2 0 1 )
Pipette 5 - 5 0 u L ( Y B 3 0 1 )
Pipette 100-1000uL(YB302)
Equipment for coliection of S a m p l e s

Sample Coliection And Preparation

U s e serum samples. Collect blood samples by venepuncture

and let them clot naturally and seprate the s e r a a s soon a s

Vers!on::;SM033E0707^

8.6

1x25.0 mL

6. Storage and Stabllity

T h e unopened reagent kit should be stored under 2-8'C and
can be used until the expiry date labeled on the kit. Do not
freeze! T h e buffer should be equilibrated to room temperature
before use. Once opened store the antisera and control at
2-8'C and the buffer at 18-25C and be s u r e to s c r e w on the
cap tightiy. Under these conditions the buffer is stable for 3
months, antisera and control for 1 month.
7.

Reagent
Sample (1/11)
C 3 Reaction Buffer
C 3 Antiserum

Kit C o m p o n e n t s
Coda
DA033

T e s t Procedure

8.8

8,9

8.10

8.11

Volume
40ul
400ul
40ul

Switch N E P H S T A R on.
Enter chemistry number. Enter chemistry number of C 3
kit (C3=33). If C 3 a s s a y h a s never been performed on
the instrument before, please swipe card when "please
swipe card" is displayed.
The a s s a y ame and lot of reagent are displayed.
C h e c k carefully, press E N T E R if the lot number is
dentical to that printed on the card or kit label, otherwise
swipe card to update the curve daa stored in
NEPHSTAR.
Dilute samples or controls using N E P H S T A R Sample
Diluent supplied in N E P H S T A R Accessory pack (Cat.
No: D K 1 1 0 ) . T h e default dilution scheme for C 3 a s s a y
is 1/11 (e.g. 4 0 0 u L sample diluent + 40uL sample) .
Prepare one cuvette for each sample to be a s s a y e d .
Place a stirring bar to the cuvette using the frceps
supplied with N E P H S T A R , then add 40uL of diluted
sample carefully to the bottom of the cuvette.
Enter sample ID. P r e s s number keys to enter the
sample ID; or press E N T E R to accept the currently
displayed sample ID.
Enter sample dilution: 1 1 . Accept the default sample
dilution by pressing E N T E R , othenwise press number
keys to alter the dilution scheme.
Place cuvette in chamber. Place the cuvette containing
a stirring bar and 4 0 u L of diluted sample in the chamber
and press it down slightiy until it reaches the bottom of
the chamber. T h e cuvette will be detected automatically.
Add reagent. Add 400 uL 0 3 reaction buffer and 40 uL
0 3 antiserum simultaneously into the cuvette using the
electronic pipette
( O a t . No.: Y B 2 0 1 ) supplied with
N E P H S T A R . N E P H S T A R will s e n s e the addition of
reagents. With movement of the stirring bar, the a s s a y
begins after blanking and result will be printed
automatically at the end of the a s s a y .
On completion of the a s s a y remove the cuvette, press
E N T E R to perform the next a s s a y . Sample ID will
increase sequentially. For alteration of the ID press
B A O K twice and tip in the right number.
If N E P H S T A R
indicates result is higher
than
measurement range, reassay the sample at a higher
dilution of e.g. 1/55 ( 1 6 0 ^ 1 sample diluent + 4 0 n L 1/1
diluted s a m p l e ) . Accordingly the sample dilution should

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