Human VEGF-D ELISA Kit - Quantikine

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DVED00
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Human VEGF-D ELISA Cell Culture Supernate Standard Curve
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Product Details
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Citations (36)
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Human VEGF-D Quantikine ELISA Kit Summary

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.

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 (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 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 (pg/mL) 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.
Human VEGF-D ELISA Linearity

Scientific Data

Human VEGF-D ELISA Cell Culture Supernate Standard Curve

Human VEGF-D ELISA Serum/Plasma Standard Curve

Knockdown Validated Human VEGF-D Quantikine ELISA Kit View Larger

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.

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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
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
&#9888; 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.

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 for Human VEGF-D Quantikine ELISA Kit

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.

36 Citations: Showing 1 - 10
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  1. Sodium accumulation in the skin is associated with higher density of skin lymphatic vessels in patients with arterial hypertension
    Authors: Chachaj, A;Stanimirova, I;Chabowski, M;Gomu?kiewicz, A;Hodurek, P;Glatzel-Pluci?ska, N;Olbromski, M;Piotrowska, A;Kuzan, A;Grzegrzó?ka, J;Ratajczak-Wielgomas, K;Nowak, A;Szahidewicz-Krupska, E;Wi?niewski, J;Bromke, MA;Podhorska-Oko?ów, M;Gamian, A;Janczak, D;Dzi?giel, P;Szuba, A;
    Advances in medical sciences
    Species: Human
    Sample Types: Plasma, Serum
  2. Early increase of plasma soluble VEGFR-2 is associated with clinical benefit from second-line treatment of paclitaxel and ramucirumab in advanced gastric cancer
    Authors: L Fornaro, G Musettini, P Orlandi, I Pecora, C Vivaldi, M Banchi, F Salani, E Fini, V Massa, S Catanese, F Cucchiara, M Lencioni, G Masi, E Vasile, G Bocci
    American journal of cancer research, 2022-07-15;12(7):3347-3356.
    Species: Human
    Sample Types: Serum
  3. Retinal VEGF-A Overexpression Is Not Sufficient to Induce Lymphangiogenesis Regardless of VEGF-C Upregulation and Lyve1+ Macrophage Infiltration
    Authors: I Wada, S Nakao, M Yamaguchi, Y Kaizu, M Arima, S Sawa, KH Sonoda
    Investigative Ophthalmology & Visual Science, 2021-10-04;62(13):17.
    Species: Human
    Sample Types: Vitreous Humor
  4. Evaluating Markers of Immune Tolerance and Angiogenesis in Maternal Blood for an Association with Risk of Pregnancy Loss
    Authors: MA Wyatt, SC Baumgarten, AL Weaver, CC Van Oort, B Fedyshyn, R Ruano, CC Shenoy, EAL Enninga
    Journal of Clinical Medicine, 2021-08-14;10(16):.
    Species: Human
    Sample Types: Plasma
  5. Neutralization of the induced VEGF-A potentiates the therapeutic effect of an anti-VEGFR2 antibody on gastric cancer in vivo
    Authors: T Mashima, T Wakatsuki, N Kawata, MK Jang, A Nagamori, H Yoshida, K Nakamura, T Migita, H Seimiya, K Yamaguchi
    Scientific Reports, 2021-07-23;11(1):15125.
    Species: Mouse
    Sample Types: Plasma
  6. Distinct Characteristics of VEGF-D and VEGF-C to Predict Mortality in Patients With Suspected or Known Coronary Artery Disease
    Authors: H Wada, M Suzuki, M Matsuda, Y Ajiro, T Shinozaki, S Sakagami, K Yonezawa, M Shimizu, J Funada, T Takenaka, Y Morita, T Nakamura, K Fujimoto, H Matsubara, T Kato, T Unoki, D Takagi, K Wada, M Wada, M Iguchi, N Masunaga, M Ishii, H Yamakage, T Kusakabe, A Yasoda, A Shimatsu, K Kotani, N Satoh-Asah, M Abe, M Akao, K Hasegawa
    J Am Heart Assoc, 2020-04-22;9(9):e015761.
    Species: Human
    Sample Types: Serum
  7. Application of non-invasive detection of peripheral vascular dysfunction in ovarian hyperstimulation syndrome (OHSS): A pilot study of clinical relevance
    Authors: TH Chin, CP Lin, SD Chang, HS Wang, HY Huang, YK Soong, HM Wu, PH Chu
    Taiwan J Obstet Gynecol, 2019-05-01;58(3):354-358.
    Species: Human
    Sample Types: Serum
  8. Serum vascular endothelial growth factor-D as a diagnostic and therapeutic biomarker for lymphangioleiomyomatosis
    Authors: M Hirose, A Matsumuro, T Arai, C Sugimoto, M Akira, M Kitaichi, LR Young, FX McCormack, Y Inoue
    PLoS ONE, 2019-02-28;14(2):e0212776.
    Species: Human
    Sample Types: Serum
  9. Dysregulation of the endothelin pathway in lymphangioleiomyomatosis with no direct effect on cell proliferation and migration
    Authors: N Chebib, F Archer, A Bobet-Erny, C Leroux, V Cottin
    Sci Rep, 2018-10-02;8(1):14698.
    Species: Human
    Sample Types: Serum
  10. Immunohistological features related to functional impairment in lymphangioleiomyomatosis
    Authors: ECTD Nascimento, BG Baldi, AW Mariani, R Annoni, RA Kairalla, SP Pimenta, LFF da Silva, CRR Carvalho, M Dolhnikoff
    Respir. Res., 2018-05-08;19(1):83.
    Species: Human
    Sample Types: Serum
  11. 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, 2016-10-04;74(1):10-21.
    Species: Human
    Sample Types: Plasma
  12. Human neural stem cell-induced endothelial morphogenesis requires autocrine/paracrine and juxtacrine signaling
    Sci Rep, 2016-07-04;6(0):29029.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. 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, 2016-01-04;11(1):e0146318.
    Species: Human
    Sample Types: Plasma
  14. 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, 2014-09-23;14(0):696.
    Species: Human
    Sample Types: Serum
  15. Filarial excretory-secretory products induce human monocytes to produce lymphangiogenic mediators.
    Authors: Weinkopff T, Mackenzie C, Eversole R, Lammie P
    PLoS Negl Trop Dis, 2014-07-10;8(7):e2893.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. 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, 2013-12-12;74(3):873-83.
    Species: Human
    Sample Types: Tissue Culture Supernates
  17. 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, 2013-02-14;61(3):755-8.
    Species: Human
    Sample Types: Serum
  18. 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, 2012-04-24;7(4):e35094.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Clinical utility of diagnostic guidelines and putative biomarkers in lymphangioleiomyomatosis.
    Respir. Res., 2012-04-18;13(0):34.
    Species: Human
    Sample Types: Serum
  20. 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, 2011-09-06;6(9):e23379.
    Species: Human
    Sample Types: Serum
  21. 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., 2011-08-01;31(8):2589-95.
    Species: Human
    Sample Types: Serum
  22. 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., 2010-10-01;163(4):776-80.
    Species: Human
    Sample Types: Serum
  23. 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, 2010-09-07;103(7):987-92.
    Species: Human
    Sample Types: Serum
  24. 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., 2010-01-22;316(6):907-14.
    Species: Human
    Sample Types: Cell Culture Supernates
  25. 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, 2009-05-01;135(5):1293-300.
    Species: Human
    Sample Types: Serum
  26. 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., 2009-04-14;284(23):16037-48.
    Species: Human
    Sample Types: Cell Culture Supernates
  27. 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., 2008-05-27;36(8):1035-46.
    Species: Human
    Sample Types: Cell Culture Supernates
  28. 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., 2008-03-01;32(3):585-92.
    Species: Human
    Sample Types: Cell Culture Supernates
  29. 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., 2007-09-18;15(12):2172-7.
    Species: Human
    Sample Types: Cell Culture Supernates
  30. 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, 2007-09-11;97(7):877-82.
    Species: Human
    Sample Types: Plasma
  31. 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., 2007-03-01;18(3):232-44.
    Species: Human
    Sample Types: Plasma, Tissue Homogenates
  32. 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., 2007-01-15;67(2):593-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  33. 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., 2006-03-01;26(2):1719-26.
    Species: Human
    Sample Types: Plasma
  34. 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., 2005-03-01;25(2):701-7.
    Species: Human
    Sample Types: Cell Culture Supernates
  35. Serum level of VEGF-D in patients with primary lymphedema.
    Authors: Fink AM, Kaltenegger I, Schneider B, Fruhauf J, Jurecka W
    Lymphology, 2004-12-01;37(4):185-9.
    Species: Human
    Sample Types: Serum
  36. 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, 2001-09-01;3(5):420-7.
    Species: Human
    Sample Types: Serum

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