Download Human Vascular Endothelial Growth Factor Receptor 2

specification sheet
Human Vascular Endothelial Growth Factor Receptor 2
Simple Plex assay for the detection of human VEGF R2 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 (pg/mL) 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.
MEAN
(pg/mL)
RANGE
(pg/mL)
STD DEV
(pg/mL)
Serum (n=5)
11,317
8968-12,737
1512
EDTA plasma (n=5)
11,207
8355-13,394
1906
Heparin Plasma (n=5)
10,842
8246-12,767
1828
SAMPLE TYPE
ULOQ
Precision
Intra-assay Precision: Each control was tested 16 times
in one assay.
LLOQ
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 VEGF R2.
CONC.
(pg/mL)
LLOQ
54.3
ULOQ
31,900
Limit of Detection
The limit of detection (LOD) of human VEGF R2 is
14.9 pg/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 (pg/mL)
222
10,677
Intra-assay SD
12.6
529
Intra-assay CV (%)
5.6
5.0
Inter-Mean (pg/mL)
193
9248
Inter-assay SD
24.3
944
Inter-assay CV (%)
12.6
10.2
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 was not necessary as VEGF R2 shows natural
linearity data from a neat sample to a 1:16 dilution.
Simple Plex Assay: Human Vascular Endothelial Growth Factor Receptor 2
specification sheet
Linearity
Specificity (continued)
Samples containing and/or spiked with high concentrations of human VEGF R2 were serially diluted with Sample
Diluent to produce samples within the dynamic range of
the assay.
The 111, 121,145, and 189 amino acid variants of VEGF-A
do not cross-react in this assay, but demonstrate a low
level of interference. Recombinant proteins were added
at the level indicated in the table to a mid-level VEGF R2
control. The VEGF R2 control value was decreased by the
indicated percentage in the Simple Plex assay.
SERUM
(n=4)
EDTA
PLASMA
(n=4)
HEPARIN
PLASMA
(n=4)
DILUTION
PARAMETER
1:2
Avg % of
Expected
96
102
102
Range (%)
88-104
88-114
95-106
Avg % of
Expected
101
105
94
Range (%)
92-109
91-127
89-99
Avg % of
Expected
89
99
96
Range (%)
82-97
81-112
84-103
Avg % of
Expected
91
97
94
Range (%)
80-101
87-110
86-102
1:4
1:8
1:16
% DECREASE
human VEGF-A111
1563
16
human VEGF-A121
3125
15
human VEGF-A145
6250
5
human VEGF-A189
3125
5
Sample Collection and Storage
The sample collection and storage conditions listed
below are intended as general guidelines. Sample
stability has not been evaluated.
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
This assay recognizes natural and recombinant human
VEGF R2. The factors listed were prepared at 50 ng/
mL in Sample Diluent and assayed for cross-reactivity.
Preparations of the following factors at 50 ng/mL in a
rhVEGF R2 control were assayed for interference. No
significant cross-reactivity or interference was observed.
Recombinant human:
• Angiogenin
• Angiopoietin-like 7
• Tie-1
• Angiopoietin-1
• Tie-2
• Angiopoietin-2
• VEGF-A165
• Angiopoietin-4
• VEGF-B167
• Angiopoietin-like 3
• VEGF-C
• Angiopoietin-like 4
• VEGF-D
• VEGF/PlGF
• Angiopoietin-like 5
• VEGF R1
• Angiopoietin-like 6
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]
proteinsimple.com
proteinsimple.com
AMOUNT ADDED
(pg/mL)
RECOMBINANT FACTOR
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.
Note: Grossly hemolyzed or icteric samples are not suitable for
use with this assay.
Sample Preparation
Serum and plasma samples require a minimum 10-fold
dilution with sample diluent. Samples above the ULOQ
require further dilution.
© 2015 ProteinSimple. Ella, Simple Plex and the
ProteinSimple logo are trademarks and/or registered
trademarks of ProteinSimple.
2
D10-1022-001 / Rev A
Rev B.