specification sheet
Human Cancer Antigen 125
Simple Plex assay for the detection of human CA125/MUC16 in
serum and plasma (EDTA/Heparin)
For research use only. Not for use in diagnostic procedures.
Calibration Curve
Endogenous Levels
The factory generated calibration curve shown below
was compiled by averaging 5 replicates of each calibrator
from multiple runs. The 5PL curve fit shows calibrator
concentration as a function of signal intensity (relative
fluorescent units, RFU).
Endogenous levels were calculated from multiple
samples. Samples were from apparently healthy
volunteers. No medical histories were available for the
donors in this study.
ULOQ
SAMPLE TYPE
MEAN
(mU/mL)
RANGE
(mU/mL)
STD DEV
(mU/mL)
Serum (n=10)
11,317
4440-22,600
4915
EDTA Plasma (n=10)
11,290
4640-21,140
4534
Heparin Plasma (n=10)
11,931
4330-23,570
5535
Precision
LLOQ
Intra-assay Precision: Each control was tested 16 times
in one assay.
Inter-assay Precision: Replicates of each control were
tested in multiple assays performed by at least three
technicians using two lots of reagents.
PARAMETER
Limit of Quantitation
Data shown represents typical performance results for
Lower Limit of Quantitation (LLOQ) and Upper Limit of
Quantitation (ULOQ) of CA125.
CONC.
(mU/mL)
LLOQ
306
ULOQ
471,920
Limit of Detection
The limit of detection (LOD) of human CA125 is
158 mU/mL. The LOD was calculated by adding three
standard deviations to the mean background signal
determined from multiple runs.
LOW QC
HIGH QC
Intra-Mean (mU/mL)
2954
141,869
Intra-assay SD
55.9
3239
Intra-assay CV (%)
1.9
2.3
Inter-Mean (mU/mL)
2885
130,993
Inter-assay SD
152
8524
Inter-assay CV (%)
5.3
6.5
Correlation
This assay has been correlated to the Quantikine® ELISA
Kit with a slope of 0.9-1.1 and an R2 value greater than 0.9.
Recovery
Recovery at three different spiked concentrations within
the range of the assay was evaluated.
AVG %
RANGE %
Serum (n=4)
SAMPLE TYPE
95
88-99
EDTA Plasma (n=4)
101
97-106
Heparin Plasma (n=4)
93
90-98
Simple Plex Assay: Human Cancer Antigen 125
specification sheet
Linearity
Calibration
Samples containing high concentrations of human
CA125 were serially diluted with Sample Diluent to
produce samples within the dynamic range of the assay.
This immunoassay is calibrated against a highly purified
CHO cell-expressed recombinant human CA125/MUC16
produced at R&D Systems. Unit values were assigned by
methods outlined in Klug, T.L. et al. (1984) Cancer Res.
44:1048.
SERUM
(n=4)
EDTA
PLASMA
(n=4)
HEPARIN
PLASMA
(n=4)
DILUTION
PARAMETER
1:2
Avg % of
Expected
108
101
105
Range (%)
105-109
96-105
101-109
Avg % of
Expected
108
103
102
Range (%)
104-112
96-108
98-109
Avg % of
Expected
104
106
107
Range (%)
74-118
102-111
104-112
Avg % of
Expected
112
113
105
Range (%)
106-119
106-120
99-113
1:4
1:8
1:16
Sample Collection and Storage
The sample collection and storage conditions listed
below are intended as general guidelines. Sample
stability has not been evaluated.
CA125 is found in saliva. Take necessary precautions
(e.g. mask and gloves) to prevent contamination of
the kit reagents while running this assay.
Serum: Use a serum separator tube (SST) and allow
samples to clot for 30 minutes at room temperature
before centrifugation for 15 minutes at 1000 x g. Remove
serum and assay immediately or aliquot and store samples at ≤ -20° C. Avoid repeated freeze-thaw cycles.
Specificity
Plasma: Collect plasma using heparin or EDTA as an
anticoagulant. Centrifuge for 15 minutes at 1000 x g
within 30 minutes of collection. Assay immediately or
aliquot and store samples at ≤ -20° C. Avoid repeated
freeze-thaw cycles.
This assay recognizes natural and recombinant
human CA125. The factors listed were prepared at
50 ng/mL in Sample Diluent and assayed for crossreactivity. Preparations of the following factors at
50 ng/mL in a rhCA125 control were assayed for
interference. No significant cross-reactivity or interference
was observed.
Recombinant human:
• CD300LG/Nepmucin
• E-Cadherin
• Endomucin
• Galectin-1
• Galectin-3
• Mesothelin
Note: Grossly hemolyzed or icteric samples are not suitable for
use with this assay.
Sample Preparation
• MMR/CD206
• MUC-1
• MUC-4
• MUC-20
• MUCDHL
Serum and plasma samples require a minimum 10-fold
dilution with sample diluent. Samples above the ULOQ
require further dilution.
Toll-free:(888)
Toll-free:
(888)607-9692
607-9692
Tel: (408)
(408)510-5500
510-5500
Fax: (408)
Fax:
(408)510-5599
510-5599
[email protected]
[email protected]
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2
D10-1038-001 / Rev A
Rev B.