Quantikine® ELISA
This package insert must be read in its entirety before using this product.
For research use only. Not for use in diagnostic procedures.
Catalog Number DVE00
Catalog Number SVE00
Catalog Number PDVE00
For the quantitative determination of human Vascular Endothelial Growth Factor （VEGF） concentrations in cell culture supernates, serum, and plasma.
MANUFACTURED AND DISTRIBUTED BY:
USA & Canada | R&D Systems, Inc.
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（800） 343-7475 （612） 379-2956 （612） 656-4400
: info@RnDSystems.com
DISTRIBUTED BY:
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+44 （0）1235 529449 +44 （0）1235 533420
: info@RnDSystems.co.uk
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+86 （21） 52380373 +86 （21） 52371001
: info@RnDSystemsChina.com.cn
TABLE OF CONTENTS
SECTION PAGE
INTRODUCTION.....................................................................................................................................................................1
PRINCIPLE OF THE ASSAY...................................................................................................................................................2
LIMITATIONS OF THE PROCEDURE.................................................................................................................................2
TECHNICAL HINTS.................................................................................................................................................................2
MATERIALS PROVIDED & STORAGE CONDITIONS...................................................................................................3
OTHER SUPPLIES REQUIRED.............................................................................................................................................4
PRECAUTIONS.........................................................................................................................................................................4
SAMPLE COLLECTION & STORAGE.................................................................................................................................4
REAGENT PREPARATION.....................................................................................................................................................5
ASSAY PROCEDURE .............................................................................................................................................................6
CALCULATION OF RESULTS...............................................................................................................................................7
TYPICAL DATA.........................................................................................................................................................................7
PRECISION................................................................................................................................................................................8
RECOVERY................................................................................................................................................................................8
SENSITIVITY.............................................................................................................................................................................8
LINEARITY.................................................................................................................................................................................9
CALIBRATION..........................................................................................................................................................................9
SAMPLE VALUES..................................................................................................................................................................10
SPECIFICITY...........................................................................................................................................................................11
REFERENCES.........................................................................................................................................................................12
PLATE LAYOUT.....................................................................................................................................................................13
www.RnDSystems.com 1
INTRODUCTION
Vascular endothelial growth factor （VEGF or VEGF-A）, also known as vascular permeability
factor （VPF）, is a potent mediator of both angiogenesis and vasculogenesis in the fetus and
adult （1-3）. It is a member of the PDGF family that is characterized by the presence of eight
conserved cysteine residues in a cystine knot structure and the formation of antiparallel
disulfide-linked dimers （4）. Humans express alternay spliced isoforms of 121, 145, 165, 183,
189, and 206 amino acids （aa） in length （4）. VEGF165 appears to be the most abundant and
potent isoform, followed by VEGF121 and VEGF189 （3, 4）. Isoforms other than VEGF121 contain
basic heparin-binding regions and are not freely diffusible （4）. Human VEGF165 shares 88% aa
sequence identity with corresponding regions of mouse and rat VEGF. VEGF is expressed in
multiple cells and tissues including skeletal and cardiac muscle （5, 6）, hepatocytes （7）,
osteoblasts （8）, neutrophils （9）, macrophages （10）, keratinocytes （11）, brown adipose tissue
（12）, CD34+ stem cells （13）, endothelial cells （14）, fibroblasts, and vascular smooth muscle cells
（15）. VEGF expression is induced by hypoxia and cytokines such as IL-1, IL-6, IL-8, oncostatin M
and TNF-α （3, 4, 9, 16）. VEGF isoforms are differentially expressed during development and in
the adult （3）.
VEGF dimers bind to two related receptor tyrosine kinases, VEGF R1 （also called Flt-1） and
VEGF R2 （Flk-1/KDR）, and induce their homodimerization and autophosphorylation （3, 4, 7, 17,
18）. These receptors have seven extracellular immunoglobulin-like domains and an intracellular
split tyrosine kinase domain. They are expressed on vascular endothelial cells and a range of
non-endothelial cells. Although VEGF affinity is highest for binding to VEGF R1, VEGF R2
appears to be the primary mediator of VEGF angiogenic activity （3, 4）. VEGF165 also binds the
semaphorin receptor, neuropilin-1, which promotes complex formation with VEGF R2 （19）.
VEGF is best known for its role in vasculogenesis. During embryogenesis, VEGF regulates the
proliferation, migration, and survival of endothelial cells （3, 4）, thus regulating blood vessel
density and size, but playing no role in determining vascular patterns. VEGF promotes bone
formation through osteoblast and chondroblast recruitment and is also a monocyte
chemoattractant （20-22）. After birth, VEGF maintains endothelial cell integrity and is a potent
mitogen for micro- and macro-vascular endothelial cells. In adults, VEGF functions mainly in
wound healing and the female reproductive cycle （3）. In diseased tissues, VEGF promotes
vascular permeability. It is thus thought to contribute to tumor metastasis by promoting both
extravasation and tumor angiogenesis （23, 24）. Various strategies have been employed
therapeutically to antagonize VEGF-mediated tumor angiogenesis （25）. Circulating VEGF levels
correlate with disease activity in autoimmune diseases such as rheumatoid arthritis, multiple
sclerosis and systemic lupus erythematosus （26）.
The Quantikine Human VEGF Immunoassay is a 4.5 hour solid phase ELISA designed to
measure VEGF165 levels in cell culture supernates, serum, and plasma. It contains
Sf 21-expressed recombinant human VEGF165 and antibodies raised against the recombinant
protein. Results obtained for naturally occurring human VEGF and recombinant human VEGF121
showed linear curves that were parallel to the standard curves obtained using the Quantikine
Human VEGF Immunoassay standards. These results indicate that this kit can be used to
determine relative mass values for natural human VEGF.
2 For research use only. Not for use in diagnostic procedures.
PRINCIPLE OF THE ASSAY
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for VEGF has been pre-coated onto a microplate. Standards and samples are
pipetted into the wells and any VEGF present is bound by the immobilized antibody. After
washing away any unbound substances, an enzyme-linked polyclonal antibody specific for
VEGF is added to the wells. Following a wash to remove any unbound antibody-enzyme
reagent, a substrate solution is added to the wells and color develops in proportion to the
amount of VEGF bound in the initial step. The color development is stopped and the intensity
of the color is measured.
LIMITATIONS OF THE PROCEDURE
• FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
• The kit should not be used beyond the expiration date on the kit label.
• Do not mix or substitute reagents with those from other lots or sources.
• It is important that the Calibrator Diluent selected for the standard curve be consistent with
the samples being assayed.
• If samples generate values higher than the highest standard, dilute the samples with the
appropriate Calibrator Diluent and repeat the assay.
• Any variation in standard diluent, operator, pipetting technique, washing technique,
incubation time or temperature, and kit age can cause variation in binding.
• Variations in sample collection, processing, and storage may cause sample value differences.
• This assay is designed to eliminate interference by soluble receptors, binding proteins,
and other factors present in biological samples. Until all factors have been tested in the
Quantikine Immunoassay, the possibility of interference cannot be excluded.
TECHNICAL HINTS
• When mixing or reconstituting protein solutions, always avoid foaming.
• To avoid cross-contamination, change pipette tips between additions of each standard level,
between sample additions, and between reagent additions. Also, use separate reservoirs for
each reagent.
• To ensure accurate results, proper adhesion of plate sealers during incubation steps is
necessary.
• When using an automated plate washer, adding a 30 second soak period following the
addition of Wash Buffer, and/or rotating the plate 180 degrees between wash steps may
improve assay precision.
• Substrate Solution should remain colorless until added to the plate. Keep Substrate Solution
protected from light. Substrate Solution should change from colorless to gradations of blue.
• Stop Solution should be added to the plate in the same order as the Substrate Solution. The
color developed in the wells will turn from blue to yellow upon addition of the Stop Solution.
Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with
the Substrate Solution.
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MATERIALS PROVIDED & STORAGE CONDITIONS
Store the unopened kit at 2-8 °C. Do not use past kit expiration date.
PART PART #
CATALOG #
DVE00
CATALOG #
SVE00 DESCRIPTION
STORAGE OF OPENED/
RECONSTITUTED MATERIAL
VEGF
Microplate
890218 1 plate 6 plates 96 well polystyrene microplate
（12 strips of 8 wells） coated with
a mouse monoclonal antibody
against VEGF.
Return unused wells to the foil
pouch containing the desiccant
pack. Reseal along entire edge of
zip-seal. May be stored for up to
1 month at 2-8 °C.*
VEGF
Standard
890220 3 vials 18 vials 2000 pg/vial of recombinant
VEGF165 in a buffered protein base
with preservatives; lyophilized.
Discard the VEGF stock solution
and dilutions after 4 hours. Use a
fresh standard for each assay.
VEGF
Conjugate
890219 1 vial 6 vials 21 mL/vial of a polyclonal
antibody against VEGF conjugated
to horseradish peroxidase with
preservatives.
May be stored for up to
1 month at 2-8 °C.*
Assay Diluent
RD1W
895117 1 vial 6 vials 11 mL/vial of a buffered protein
base with preservatives.
Calibrator Diluent
RD5K
895119 1 vial 6 vials 21 mL/vial of a buffered protein
base with preservatives. For cell
culture supernate samples.
Calibrator Diluent
RD6U
895148 1 vial 6 vials 21 mL/vial of animal serum with
preservatives. For serum/plasma
samples.
Wash Buffer
Concentrate
895003 1 vial 6 vials 21 mL/vial of a 25-fold
concentrated solution of buffered
surfactant with preservatives.
Color Reagent A 895000 1 vial 6 vials 12 mL/vial of stabilized hydrogen
peroxide.
Color Reagent B 895001 1 vial 6 vials 12 mL/vial of stabilized chromogen
（tetramethylbenzidine）.
Stop Solution 895032 1 vial 6 vials 6 mL/vial of 2 N sulfuric acid.
Plate Sealers N/A 4 strips 24 strips Adhesive strips.
* Provided this is within the expiration date of the kit.
DVE00 contains sufficient materials to run an ELISA on one 96 well plate.
SVE00 （SixPak） contains sufficient materials to run ELISAs on six 96 well plates.
This kit is also available in a PharmPak （R&D Systems, Catalog # PDVE00）. PharmPaks contain
sufficient materials to run ELISAs on 50 microplates. Specific vial counts of each component
may vary. Please refer to the literature accompanying your order for specific vial counts.
4 For research use only. Not for use in diagnostic procedures.
OTHER SUPPLIES REQUIRED
• Microplate reader capable of measuring absorbance at 450 nm, with the correction
wavelength set at 540 nm or 570 nm.
• Pipettes and pipette tips.
• Deionized or distilled water.
• Squirt bottle, manifold dispenser, or automated microplate washer.
• 500 mL graduated cylinder.
• Polypropylene test tubes for dilution of standards.
• Human VEGF Controls （optional; available from R&D Systems）.
PRECAUTIONS
Calibrator Diluent RD6U contains sodium azide which may react with lead and copper
plumbing to form explosive metallic azides. Flush with large volumes of water during disposal.
VEGF is detectable in saliva. Take precautionary measures to prevent contamination of the kit
reagents while running the assay.
The Stop Solution provided with this kit is an acid solution. Wear protective gloves, clothing,
eye, and face protection. Wash hands thoroughly after handling.
SAMPLE COLLECTION & STORAGE
The sample collection and storage conditions listed below are intended as general
guidelines. Sample stability has not been evaluated.
Cell Culture Supernates - Cell culture supernates should contain at least 1% fetal calf serum
for stability of the VEGF. Remove particulates by centrifugation and assay immediay or
aliquot and store samples at ≤ -20 °C. Avoid repeated freeze-thaw cycles.
Serum - Use a serum separator tube （SST） and allow samples to clot for 30 minutes before
centrifugation for 15 minutes at 1000 x g. Remove serum and assay immediay or aliquot and
store samples at ≤ -20 °C. Avoid repeated freeze-thaw cycles.
Plasma - Collect plasma using EDTA, heparin, or citrate as an anticoagulant. Centrifuge for
15 minutes at 1000 x g within 30 minutes of collection. Assay immediay or aliquot and store
samples at ≤ -20 °C. Avoid repeated freeze-thaw cycles.
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REAGENT PREPARATION
Bring all reagents to room temperature before use.
Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mix
gently until the crystals have compley dissolved. Dilute 20 mL of Wash Buffer Concentrate
into deionized or distilled water to prepare 500 mL of Wash Buffer.
Substrate Solution - Color Reagents A and B should be mixed together in equal volumes
within 15 minutes of use. Protect from light. 200 μL of the resultant mixture is required per well.
VEGF Standard - Reconstitute the VEGF Standard with 1.0 mL of Calibrator Diluent RD5K （for
cell culture supernate samples） or Calibrator Diluent RD6U （for serum/plasma samples）. This
reconstitution produces a stock solution of 2000 pg/mL. Allow the standard to sit for a
minimum of 15 minutes with gentle agitation prior to making dilutions.
For Cell Culture Supernate Samples: Use polypropylene tubes. Pipette 500 μL of Calibrator
Diluent RD5K into each tube. Use the stock solution to produce a dilution series （below）. Mix
each tube thoroughly before the next transfer. The 1000 pg/mL dilution serves as the high
standard. Calibrator Diluent RD5K serves as the zero standard （0 pg/mL）.
500 μL Std.
2000 pg/mL 1000 pg/mL 500 pg/mL 250 pg/mL 125 pg/mL 62.5 pg/mL 31.2 pg/mL 15.6 pg/mL
500 μL 500 μL 500 μL 500 μL 500 μL 500 μL
500 μL Std.
2000 pg/mL 1000 pg/mL 500 pg/mL 250 pg/mL 125 pg/mL 62.5 pg/mL 31.2 pg/mL
500 μL 500 μL 500 μL 500 μL 500 μL
For Serum/Plasma Samples: Use polypropylene tubes. Pipette 500 μL of Calibrator Diluent
RD6U into each tube. Use the stock solution to produce a dilution series （below）. Mix each
tube thoroughly before the next transfer. The undiluted standard serves as the high standard
（2000 pg/mL）. Calibrator Diluent RD6U serves as the zero standard （0 pg/mL）
6 For research use only. Not for use in diagnostic procedures.
ASSAY PROCEDURE
Bring all reagents and samples to room temperature before use. It is recommended that
all standards, samples, and controls be assayed in duplicate.
1. Prepare all reagents, working standards, and samples as directed in the previous sections.
2. Remove excess microplate strips from the plate frame, return them to the foil pouch
containing the desiccant pack, and reseal.
3. For Cell Culture Supernate Samples: Add 50 μL of Assay Diluent RD1W to each well.
For Serum/Plasma Samples: Add 100 μL of Assay Diluent RD1W to each well.
4. For Cell Culture Supernate Samples: Add 200 μL of Standard, control, or sample per well.
For Serum/Plasma Samples: Add 100 μL of Standard, control, or sample per well.
Cover with the adhesive strip provided and incubate for 2 hours at room temperature.
A plate layout is provided to record the standards and samples assayed.
5. Aspirate each well and wash, repeating the process twice for a total of three washes. Wash
by filling each well with Wash Buffer （400 μL） using a squirt bottle, manifold dispenser, or
autowasher. Complete removal of liquid at each step is essential to good performance.
After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert
the plate and blot it against clean paper towels.
6. Add 200 μL of VEGF Conjugate to each well. Cover with a new adhesive strip. Incubate for
2 hours at room temperature.
7. Repeat the aspiration/wash as in step 5.
8. Add 200 μL of Substrate Solution to each well. Protect from light.
For Cell Culture Supernate Samples: Incubate for 20 minutes at room temperature.
For Serum/Plasma Samples: Incubate for 25 minutes at room temperature.
9. Add 50 μL of Stop Solution to each well. If color change does not appear uniform, gently
tap the plate to ensure thorough mixing. If the color in the wells is green or the color
change does not appear uniform, gently tap the plate to ensure thorough mixing.
10. Determine the optical density of each well within 30 minutes, using a microplate reader
set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength
correction is not available, subtract readings at 540 nm or 570 nm from the readings at
450 nm. This subtraction will correct for optical imperfections in the plate. Readings made
directly at 450 nm without correction may be higher and less accurate.
www.RnDSystems.com 7
CALCULATION OF RESULTS
Average the duplicate readings for each standard, control, and sample and subtract the
average zero standard optical density.
Create a standard curve by reducing the data using computer software capable of generating a
four parameter logistic （4-PL） curve fit. As an alternative, construct a standard curve by plotting
the mean absorbance for each standard on the y-axis against the concentration on the x-axis
and draw a best fit curve through the points on the graph. The data may be linearized by
plotting the log of the VEGF concentrations versus the log of the O.D. and the best fit line can
be determined by regression analysis. This procedure will produce an adequate but less precise
fit of the data.
If samples have been diluted, the concentration read from the standard curve must be
multiplied by the dilution factor.
TYPICAL DATA
These standard curves are provided for demonstration only. A standard curve should be
generated for each set of samples assayed.
（pg/mL） O.D. Average Corrected
0 0.074 0.075 —
0.076
15.6 0.118 0.120 0.045
0.121
31.2 0.159 0.159 0.084
0.159
62.5 0.246 0.244 0.169
0.242
125 0.384 0.381 0.306
0.378
250 0.666 0.668 0.593
0.669
500 1.258 1.260 1.185
1.263
1000 2.302 2.268 2.193
2.233
（pg/mL） O.D. Average Corrected
0 0.068 0.070 —
0.071
31.2 0.107 0.108 0.038
0.110
62.5 0.149 0.151 0.081
0.153
125 0.230 0.230 0.160
0.230
250 0.377 0.382 0.312
0.387
500 0.657 0.678 0.608
0.699
1000 1.261 1.271 1.201
1.281
2000 2.159 2.202 2.132
2.246
CALIBRATOR DILUENT RD5K
CALIBRATOR DILUENT RD6U
8 For research use only. Not for use in diagnostic procedures.
PRECISION
Intra-assay Precision （Precision within an assay）
Three samples of known concentration were tested twenty times on one plate to assess intraassay
precision.
Inter-assay Precision （Precision between assays）
Three samples of known concentration were tested in forty separate assays to assess interassay
precision.
CELL CULTURE SUPERNATE ASSAY
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean （pg/mL） 29.1 123 531 32.8 128 495
Standard deviation 1.9 5.0 18.4 2.8 6.4 33.0
CV （%） 6.5 4.1 3.5 8.5 5.0 6.7
SERUM/PLASMA ASSAY
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean （pg/mL） 53.7 235 910 64.5 250 1003
Standard deviation 3.6 10.6 46.2 5.7 17.4 61.7
CV （%） 6.7 4.5 5.1 8.8 7.0 6.2
RECOVERY
The recovery of VEGF spiked to three different levels throughout the range of the assay in
various matrices was evaluated.
Sample Type Average % Recovery Range
Cell culture media （n=5） 102 95-111%
Serum （n=5） 102 92-115%
EDTA plasma （n=5） 97 82-113%
Heparin plasma （n=5） 93 82-102%
Citrate plasma （n=5） 100 88-113%
SENSITIVITY
Using Calibrator Diluent RD5K the minimum detectable dose （MDD） of VEGF is typically less
than 5.0 pg/mL. Using Calibrator Diluent RD6U the MDD is typically less than 9.0 pg/mL.
The MDD was determined by adding two standard deviations to the mean optical density
value of twenty zero standard replicates and calculating the corresponding concentration.
www.RnDSystems.com 9
LINEARITY
To assess linearity of the assay, samples were spiked with high concentrations of VEGF and
diluted with the appropriate Calibrator Diluent to produce samples with values within the
dynamic range of the assay.
Cell culture
media
（n=5）
Serum
（n=5）
EDTA
plasma
（n=5）
Heparin
plasma
（n=5）
Citrate
plasma
（n=5）
1:2
Average % of Expected 98 97 97 94 95
Range （%） 94-100 91-103 82-107 87-99 90-100
1:4
Average % of Expected 96 97 98 93 94
Range （%） 93-99 93-104 91-106 85-98 89-99
1:8
Average % of Expected 93 96 96 92 92
Range （%） 88-102 93-103 89-106 85-101 85-97
1:16
Average % of Expected 93 94 94 94 92
Range （%） 88-105 91-101 84-106 83-103 85-98
CALIBRATION
This immunoassay is calibrated against a highly purified Sf 21-expressed recombinant human
VEGF165 produced at R&D Systems.
The NIBSC/WHO VEGF165 preparation 02/286 （recombinant human DNA） was evaluated in this
kit. The dose response curve of the standard 02/286 parallels the Quantikine standard curve.
To convert sample values obtained with the Quantikine Human VEGF kit to approximate
NIBSC/WHO 02/286 Units, use the equation below.
NIBSC/WHO （02/286） approximate value （U/mL） = 0.002 x Quantikine VEGF value （pg/mL）
Note: Based on data generated in April 2011.
10 For research use only. Not for use in diagnostic procedures.
SAMPLE VALUES
Serum/Plasma - Samples from apparently healthy volunteers were evaluated for the presence
of VEGF in this assay. No medical histories were available for the donors used in this study.
Sample Type Mean of Detectable （pg/mL） % Detectable Range （pg/mL）
Serum （n=37） 220 100 62-707
EDTA plasma （n=37） 61 24 ND-115
Heparin plasma （n=37） 41 22 ND-55
Citrate plasma （n=37） ___ 0 ND
ND=Non-detectable
Cell Culture Supernates - Human peripheral blood mononuclear cells （1 x 106 cells/mL）
were cultured in RPMI supplemented with 5% fetal calf serum, 50 μM β-mercaptoethanol,
2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were
cultured unstimulated or stimulated with 10 μg/mL PHA for 1 and 5 days. Aliquots of the cell
culture supernates were removed and assayed for levels of natural VEGF.
Condition Day 1 （pg/mL） Day 5 （pg/mL）
Unstimulated 356 332
Stimulated 14 1440
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SPECIFICITY
This assay recognizes natural and recombinant human VEGF. This assay also recognizes
recombinant human VEGF165b.
The factors listed below were prepared at 50 ng/mL in Calibrator Diluent and assayed for crossreactivity.
Preparations of the following factors at 50 ng/mL in a mid-range VEGF control were
assayed for interference. The following factors showed no cross-reactivity or interference.
Recombinant human:
PDGF-AA
PDGF-AB
PDGF-BB
PDGF-CC
PDGF-DD
PlGF
PlGF-2
VEGF165/PlGF
VEGF-B167
VEGF-C
VEGF-D
VEGF R3
Recombinant mouse:
PDGF-CC
PlGF-2
VEGF120
VEGF164
VEGF R3
Recombinant rat:
PDGF-AA
PDGF-AB
PDGF-BB
VEGF164
Recombinant zebrafish:
VEGF
Natural proteins:
human PDGF
porcine PDGF
VEGF-related factors showing cross-reactivity or interference.
Recombinant human VEGF R1/Flt-1 Interference at levels ≥ 500 pg/mL
Recombinant human VEGF R2/KDR Interference at levels ≥ 2000 pg/mL
Recombinant mouse VEGF R1/Flk-1 Interference at levels ≥ 500 pg/mL
Recombinant mouse VEGF R2/KDR Interference at levels ≥ 4000 pg/mL
Recombinant canine VEGF Cross-reacts approximay 67%
Recombinant feline VEGF Cross-reacts approximay 82%
12 For research use only. Not for use in diagnostic procedures.
REFERENCES
1. Leung, D.W. et al. （1989） Science 246:1306.
2. Keck, P.J. et al. （1989） Science 246:1309.
3. Byrne, A.M. et al. （2005） J. Cell. Mol. Med. 9:777.
4. Robinson, C.J. and S.E. Stringer （2001） J. Cell. Sci. 114:853.
5. Richardson, R.S. et al. （1999） Am. J. Physiol. 277:H2247.
6. Sugishita, Y. et al. （2000） Biochem. Biophys. Res. Commun. 268:657.
7. Yamane, A. et al. （1994） Oncogene 9:2683.
8. Goad, D.L. et al. （1996） Endocrinology 137:2262.
9. Gaudry, M. et al. （1997） Blood 90:4153.
10. Mclaren, J. et al. （1996） J. Clin. Invest. 98:482.
11. Diaz, B.V. et al. （2000） J. Biol. Chem. 275:642.
12. Asano, A. et al. （1997） Biochem. J. 328:179.
13. Bautz, F. et al. （2000） Exp. Hematol. 28:700.
14. Namiki, A. et al. （1995） J. Biol. Chem. 270:31189.
15. Nauck, M. et al. （1997） Am. J. Respir. Cell. Mol. Biol. 16:398.
16. Angelo, L.S. and R. Kurzrock （2007） Clin. Cancer Res. 13:2825.
17. Neufeld, G. et al. （1999） FASEB. J. 13:9.
18. Kowalewski, M.P. et al. （2005） Accession #ABB82619.
19. Pan, Q. et al. （2007） J. Biol. Chem. 282:24049.
20. Dai, J. and A.B. Rabie （2007） J. Dent. Res. 86:937.
21. Breier, G. （2000） Semin. Thromb. Hemost. 26:553.
22. Barleon, B. et al. （1996） Blood 87:3336.
23. Weis, S.M. and D.A. Cheresh （2005） Nature 437:497.
24. Thurston, G. （2002） J. Anat. 200:575.
25. Grothey, A. and E. Galanis （2009） Nat. Rev. Clin. Oncol. 6:507.
26. Carvalho, J.F. et al. （2007） J. Clin. Immunol. 27:246.
www.RnDSystems.com 13
PLATE LAYOUT
Use this plate layout to record standards and samples assayed.
14 For research use only. Not for use in diagnostic procedures.