Human VEGF-D Quantikine ELISA Kit

  (26 citations)     
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Assay Procedure
Citations (26)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
  • Sensitivity
    31.3 pg/mL
  • Assay Range
    125.0 - 4,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • 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.
Control Available
QC20 , Quantikine Immunoassay Control Group 3 - Please Inquire
Product Summary
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.

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 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 7.4 5.9

Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 448 970 2081 474 956 2013
Standard Deviation 27.8 40.9 49.3 38 68.5 145
CV% 6.2 4.2 2.4 8 7.2 7.2

Recovery

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.
 VEGF-D [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: VEGF-D

Vascular endothelia growth factor D (VEGF-D), also known as c-fos-induced growth factor (FIGF), is a secreted glycoprotein of the VEGF/PDGF family. VEGFs regulate angiogenesis and lymphangiogenesis during development and tumor growth, and are characterized by eight conserved cysteine residues that form a cystine knot structure. VEGF-C and VEGF-D, which share 23% amino acid (aa) sequence identity, are uniquely expressed as preproproteins that contain long N- and C-terminal propeptide extensions around the VEGF homology domain (VHD). Proteolytic processing of the 354 aa VEGF-D preproprotein creates a secreted proprotein. Further processing by extracellular serine proteases, such as plasmin or furin-like proprotein convertases, forms mature VEGF-D consisting of non-covalently linked 42 kDa homodimers of the 117 aa VHD.

Mature human VEGF-D shares 94%, 95%, 99%, 97% and 93% aa identity with mouse, rat, equine, canine and bovine VEGF-D, respectively. It is expressed in adult lung, heart, muscle, and small intestine, and is most abundantly expressed in fetal lungs and skin. Mouse and human VEGF-D are ligands for VEGF receptor 3 (VEGF R3, also called Flt-4) that are active across species and show enhanced affinity when processed. Processed human VEGF-D is also a ligand for VEGF R2, also called Flk-1 or KDR. 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, VEGF-D is most active in neonatal lymphatic maturation and bone growth. Both promote tumor lymphangiogenesis. Consonant with their activity on VEGF receptors, binding of VEGF-C and VEGF-D to neuropilins contributes to VEGF R3 signaling in lymphangiogenesis, while binding to integrin α9β1 mediates endothelial cell adhesion and migration.

  • Long Name:
    Vascular Endothelial Growth Factor D/cFos-induced Growth Factor
  • Entrez Gene IDs:
    2277 (Human); 14205 (Mouse)
  • Alternate Names:
    c-fos induced growth factor (vascular endothelial growth factor D); FIGF; vascular endothelial growth factor D; VEGFD; VEGF-D; VEGF-DVEGFDc-Fos-induced growth factor
Related Research Areas
Assay Procedure
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
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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.
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

26 Citations: Showing 1 - 10
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Species
Sample Type
  1. Tuberculosis-diabetes co-morbidity is characterized by heightened systemic levels of circulating angiogenic factors.
    Authors: Kumar N, Moideen K, Sivakumar S, Menon P, Viswanathan V, Kornfeld H, Babu S
    J Infect, 2017;74(1):10-21.
    Species: Human
    Sample Type: Plasma
  2. Circulating Angiogenic Factors as Biomarkers of Disease Severity and Bacterial Burden in Pulmonary Tuberculosis.
    Authors: Kumar N, Banurekha V, Nair D, Babu S
    PLoS ONE, 2016;11(1):e0146318.
    Species: Human
    Sample Type: Plasma
  3. Human neural stem cell-induced endothelial morphogenesis requires autocrine/paracrine and juxtacrine signaling
    Sci Rep, 2016;6(0):29029.
    Species: Human
    Sample Type: Cell Culture Supernates
  4. Filarial excretory-secretory products induce human monocytes to produce lymphangiogenic mediators.
    Authors: Weinkopff T, Mackenzie C, Eversole R, Lammie P
    PLoS Negl Trop Dis, 2014;8(7):e2893.
    Species: Human
    Sample Type: Cell Culture Supernates
  5. The CXCL7/CXCR1/2 axis is a key driver in the growth of clear cell renal cell carcinoma.
    Authors: Grepin R, Guyot M, Giuliano S, Boncompagni M, Ambrosetti D, Chamorey E, Scoazec J, Negrier S, Simonnet H, Pages G
    Cancer Res, 2014;74(3):873-83.
    Species: Human
    Sample Type: Tissue Culture Supernates
  6. Serum angiopoietin-2 and soluble VEGFR-2 levels predict malignancy of ovarian neoplasm and poor prognosis in epithelial ovarian cancer.
    Authors: Sallinen H, Heikura T, Koponen J, Kosma V, Heinonen S, Yla-Herttuala S, Anttila M
    BMC Cancer, 2014;14(0):696.
    Species: Human
    Sample Type: Serum
  7. Role of serum concentration of VEGFR1 and TIMP2 on clinical outcome in primary cervical cancer: results of a companion protocol of the randomized, NOGGO-AGO phase III adjuvant trial of simultaneous cisplatin-based radiochemotherapy vs. carboplatin and paclitaxel containing sequential radiotherapy.
    Authors: Braicu E, Fotopoulou C, Chekerov R, Richter R, Blohmer J, Kummel S, Stamatian F, Yalcinkaya I, Mentze M, Lichtenegger W, Sehouli J
    Cytokine, 2013;61(3):755-8.
    Species: Human
    Sample Type: Serum
  8. Co-expression of alpha9beta1 integrin and VEGF-D confers lymphatic metastatic ability to a human breast cancer cell line MDA-MB-468LN.
    Authors: Majumder M, Tutunea-Fatan E, Xin X, Rodriguez-Torres M, Torres-Garcia J, Wiebe R, Timoshenko AV, Bhattacharjee RN, Chambers AF, Lala PK
    PLoS ONE, 2012;7(4):e35094.
    Species: Human
    Sample Type: Cell Culture Supernates
  9. Clinical utility of diagnostic guidelines and putative biomarkers in lymphangioleiomyomatosis.
    Respir. Res., 2012;13(0):34.
    Species: Human
    Sample Type: Serum
  10. Prognostic significance of angiogenic factors in uterine cervical cancer.
    Authors: Landt S, Wehling M, Heidecke H, Jeschke S, Korlach S, Stoblen F, Schmid P, Blohmer JU, Lichtenegger W, Sehouli J, Kummel S
    Anticancer Res., 2011;31(8):2589-95.
    Species: Human
    Sample Type: Serum
  11. Multicenter phase 2 trial of sirolimus for tuberous sclerosis: kidney angiomyolipomas and other tumors regress and VEGF- D levels decrease.
    Authors: Dabora SL, Franz DN, Ashwal S, Sagalowsky A, DiMario FJ, Miles D, Cutler D, Krueger D, Uppot RN, Rabenou R, Camposano S, Paolini J, Fennessy F, Lee N, Woodrum C, Manola J, Garber J, Thiele EA
    PLoS ONE, 2011;6(9):e23379.
    Species: Human
    Sample Type: Serum
  12. Impaired lymphangiogenesis due to excess vascular endothelial growth factor-D/Flt-4 signalling in the skin of patients with systemic sclerosis.
    Authors: Honda N, Jinnin M, Kajihara I, Makino T, Fukushima S, Ihn H
    Br. J. Dermatol., 2010;163(4):776-80.
    Species: Human
    Sample Type: Serum
  13. Phase I and pharmacological study of the broad-spectrum tyrosine kinase inhibitor JNJ-26483327 in patients with advanced solid tumours.
    Authors: Konings IR, de Jonge MJ, Burger H
    Br. J. Cancer, 2010;103(7):987-92.
    Species: Human
    Sample Type: Serum
  14. Autocrine effects of VEGF-D on endothelial cells after transduction with AD-VEGF-D(DeltaNDeltaC).
    Authors: Papiewska-Pajak I, Boncela J, Przygodzka P, Cierniewski CS
    Exp. Cell Res., 2010;316(6):907-14.
    Species: Human
    Sample Type: Cell Culture Supernates
  15. Serum vascular endothelial growth factor-D levels in patients with lymphangioleiomyomatosis reflect lymphatic involvement.
    Authors: Glasgow CG, Avila NA, Lin JP, Stylianou MP, Moss J
    Chest, 2009;135(5):1293-300.
    Species: Human
    Sample Type: Serum
  16. Novel vascular endothelial growth factor D variants with increased biological activity.
    Authors: Toivanen PI, Nieminen T, Viitanen L, Alitalo A, Roschier M, Jauhiainen S, Markkanen JE, Laitinen OH, Airenne TT, Salminen TA, Johnson MS, Airenne KJ, Yla-Herttuala S
    J. Biol. Chem., 2009;284(23):16037-48.
    Species: Human
    Sample Type: Cell Culture Supernates
  17. Hypoxia-induced epithelial VEGF-C/VEGFR-3 upregulation in carcinoma cell lines.
    Authors: Simiantonaki N, Jayasinghe C, Michel-Schmidt R, Peters K, Hermanns MI, Kirkpatrick CJ
    Int. J. Oncol., 2008;32(3):585-92.
    Species: Human
    Sample Type: Cell Culture Supernates
  18. Isolation and characterization of CD146+ multipotent mesenchymal stromal cells.
    Authors: Sorrentino A, Ferracin M, Castelli G, Biffoni M, Tomaselli G, Baiocchi M, Fatica A, Negrini M, Peschle C, Valtieri M
    Exp. Hematol., 2008;36(8):1035-46.
    Species: Human
    Sample Type: Cell Culture Supernates
  19. Adenovirus-mediated gene transfer of human vascular endothelial growth factor-d induces transient angiogenic effects in mouse hind limb muscle.
    Authors: Kholova I, Koota S, Kaskenpaa N, Leppanen P, Narvainen J, Kavec M, Rissanen TT, Hazes T, Korpisalo P, Grohn O, Yla-Herttuala S
    Hum. Gene Ther., 2007;18(3):232-44.
    Species: Human
    Sample Type: Plasma
  20. Marrow angiogenesis-associated factors as prognostic biomarkers in patients with acute myelogenous leukaemia.
    Authors: Lee CY, Tien HF, Hu CY, Chou WC, Lin LI
    Br. J. Cancer, 2007;97(7):877-82.
    Species: Human
    Sample Type: Plasma
  21. Vascular endothelial growth factor receptor 3 is involved in tumor angiogenesis and growth.
    Authors: Laakkonen P, Waltari M, Holopainen T, Takahashi T, Pytowski B, Steiner P, Hicklin D, Persaud K, Tonra JR, Witte L, Alitalo K
    Cancer Res., 2007;67(2):593-9.
    Species: Human
    Sample Type: Cell Culture Supernates
  22. Umbilical Cord Blood-derived Progenitor Cells Enhance Muscle Regeneration in Mouse Hindlimb Ischemia Model.
    Authors: Koponen JK, Kekarainen T, E Heinonen S, Laitinen A, Nystedt J, Laine J, Yla-Herttuala S
    Mol. Ther., 2007;15(12):2172-7.
    Species: Human
    Sample Type: Cell Culture Supernates
  23. Changes in the circulating plasma levels of VEGF and VEGF-D after adjuvant chemotherapy in patients with breast cancer and 1 to 3 positive lymph nodes.
    Authors: Kummel S, Eggemann H, Luftner D, Thomas A, Jeschke S, Zerfel N, Heilmann V, Emons G, Zeiser T, Ulm K, Kobl M, Korlach S, Schmid P, Sehouli J, Elling D, Blohmer JU
    Anticancer Res., 2006;26(2):1719-26.
    Species: Human
    Sample Type: Plasma
  24. A potential role for vascular endothelial growth factor-D as an autocrine growth factor for human breast carcinoma cells.
    Authors: Akahane M, Akahane T, Shah A, Okajima E, Thorgeirsson UP
    Anticancer Res., 2005;25(2):701-7.
    Species: Human
    Sample Type: Cell Culture Supernates
  25. Serum level of VEGF-D in patients with primary lymphedema.
    Authors: Fink AM, Kaltenegger I, Schneider B, Fruhauf J, Jurecka W
    Lymphology, 2004;37(4):185-9.
    Species: Human
    Sample Type: Serum
  26. VEGF-A, VEGF-C, and VEGF-D in colorectal cancer progression.
    Authors: George ML, Tutton MG, Janssen F, Arnaout A, Abulafi AM, Eccles SA, Swift RI
    Neoplasia, 2001;3(5):420-7.
    Species: Human
    Sample Type: Serum
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