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Human VEGF-D Quantikine ELISA Kit

R&D Systems | Catalog # DVED00

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)

Sensitivity

31.3 pg/mL

Assay Range

125-4000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Human VEGF-D Quantikine ELISA Kit

The Quantikine Human VEGF-D Immunoassay is a 4.5 hour solid phase ELISA designed to measure VEGF-D levels in cell culture supernates, serum, and plasma. It contains Sf 21-expressed, recombinant human VEGF-D and antibodies raised against the recombinant protein. Results obtained for naturally occurring human VEGF-D showed linear curves that were parallel to the standard curves obtained using the kit standards. These results indicate that this kit can be used to determine relative mass values for natural human VEGF-D.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

Specificity

Natural and recombinant human VEGF-D

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 560 1098 2192 512 978 1978
Standard Deviation 31.8 35.5 45.6 46.3 72.8 117
CV% 5.7 3.2 2.1 9.0 7.4 5.9

EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 448 970 2081 474 956 2013
Standard Deviation 27.8 40.9 49.3 38.0 68.5 145
CV% 6.2 4.2 2.4 8.0 7.2 7.2

Recovery for Human VEGF-D Quantikine ELISA Kit

The recovery of VEGF-D 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=4) 100 97-104
EDTA Plasma (n=5) 105 89-114
Heparin Plasma (n=5) 104 88-114
Serum (n=5) 106 94-113

Linearity

To assess the linearity of the assay, samples were spiked with high concentrations of VEGF-D in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human VEGF-D ELISA Linearity

Scientific Data Images for Human VEGF-D Quantikine ELISA Kit

Human VEGF-D ELISA Cell Culture Supernate Standard Curve

Human VEGF-D ELISA Cell Culture Supernate Standard Curve

Human VEGF-D ELISA Serum/Plasma Standard Curve

Human VEGF-D ELISA Serum/Plasma Standard Curve

Human VEGF-D Quantikine ELISA Kit

Human VEGF-D Quantikine ELISA Kit

Reduction of migration and invasion of 468LN cells by knocking down VEGF-D production:(A–C) Levels of VEGF-D knock down in 468LN cells was confirmed by three different methods, (A) qRT-PCR, (B) western blot and (C) ELISA. After VEGF-D knock down (KD) in 468LN cells, (D) migration, invasion and (E) proliferation significantly dropped as compared to scrambled knock down (SCR-KD) cells. (F) Addition of exogenous rVEGF-D or FBS increased both migration and invasion of VEGF-D knocked down 468LN cells. Bars represent mean (n = 4 in all cases except C, n = 6) ± SE, *,P<0.05; **,P<0.01. Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0035094), licensed under a CC-BY license. Not internally tested by R&D Systems.

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: VEGF-D

Vascular endothelial growth factor D (VEGF-D) is a homodimeric ligand of VEGF R3/Flt-4 and is synthesized with N- and C-terminal propeptides. Fully processed VEGF-D containing only the VEGF homology domain can additionally bind and activate VEGF R2/KDR/Flk-1. VEGF-D is expressed in adult lung, heart, muscle, and small intestine, and is most abundantly expressed in fetal lungs and skin. VEGF R3 is strongly expressed in lymphatic endothelial cells and is essential for regulation of the growth and differentiation of lymphatic endothelium. While VEGF-C is the critical ligand for VEGF R3 during embryonic lymphatic development, while VEGF-D is most active in neonatal lymphatic maturation and bone growth.

Long Name

Vascular Endothelial Growth Factor D/cFos-induced Growth Factor

Alternate Names

FIGF, VEGFD

Entrez Gene IDs

2277 (Human); 14205 (Mouse)

Gene Symbol

VEGFD

Additional VEGF-D Products

Product Documents for Human VEGF-D Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human VEGF-D Quantikine ELISA Kit

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Human VEGF-D Quantikine ELISA Kit

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Protocols

View specific protocols for Human VEGF-D Quantikine ELISA Kit (DVED00):

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 100 µL Assay Diluent
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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FAQs

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