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Human VEGF165

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Cat-Nr.300-035S
Size2 µg
Price80 €
SourceInsect cells
Formulationlyophilized
Purity Confirmation> 95% by SDS-PAGE
Length [aa]165
Molecular Weight45.0 kDa
N Terminal SequenceAPMAEGG
Biological ActivityThe ED50 for stimulation of cell proliferation by human umbilical vein endothelial cells for VEGF165 has been determined to be in the range of 1-4 ng/ml.
Species ReactivityHuman
Buffer50mM acetic acid
ReconstitutionThe lyophilized VEGF165 should be reconstituted in 50 mM acetic acid to a concentration not lower than 50 µg/ml. For long term storage we recommend to add at least 0.1% human or bovine serum albumin.
Stability and StorageLyophilized samples are stable for greater than six months at -20°C to -70°C. Reconstituted VEGF165 should be stored in working aliquots at -20°C.
Synonymsvascular endothelial growth factor A; VEGFA; VPF; VEGF; MVCD1
DescriptionHuman Vascular Endothelial Growth Factor VEGF165, a 23 kDa protein consisting of 165 amino acid residues, is produced as a homodimer. VEGF is a polypeptide growth factor and a member of the platelet-derived growth factor family. It is a specific mitogen for vascular endothelial cells and a strong angiogenic factor in vivo. Two high-affinity tyrosine kinase receptors for VEGF165 have been identified, VEGFR-1 (FLT-1), and VEGFR-2 (KDR). Consistent with the endothelial cell-specific action of VEGF165, expression of both receptor genes has been found predominantly but not exclusively on endothelial cells. Expression of VEGFR-1 was also found on human monocytes, neutrophils (PMNs), bovine brain pericytes and villous and extra villous trophoblast. In addition to its action as a mitogen it is a potent vascular permeability factor (VPF) in vivo. VEGF165 is also a chemo attractant molecule for monocytes and endothelial cells. 5 different proteins are generated by differential splicing: VEGF121, VEGF145, VEGF165, VEGF189 and VEGF206. The most abundant form is VEGF165. Whereas VEGF121 and VEGF165 are secreted proteins, VEGF145, VEGF189 and VEGF206 are strongly cell-associated. The isoforms VEGF145, VEGF165 and VEGF189 bind to heparin with high affinity. VEGF165 is apparently a homo-dimer, but preparations of VEGF165 show some heterogeneity on SDS gels, depending on the secretion of different glycosylation patterns. All dimeric forms have similar biological activities but their bioavailability is very different. There is good evidence that different cells and tissues express different VEGF isoforms. The other members of this increasing growth factor family are VEGF-B, -C, -D and -E. Another member is the Placenta growth factor PlGF
Protein SequenceAPMAEGGGQNHHEVVKFMDVYQRSYCHPIETLVDIFQEYPDEIEYIFKPSCVPLMRCGGCCNDEGLECVPTEESNITMQIMRIKPHQGQHIGEMSFLQHNKCECRPKKDRARQENPCGPCSERRKHLFVQDPQTCKCSCKNTDSRCKARQLELNERTCRCDKPRR
Uniprot IDP15692
Protein RefSeqNP_001020537.2
mRNA RefSeqNM_001025366.2

Figures

Figure 1.
SDS-PAGE analysis of recombinant human VEGF165. Sample was loaded in 15% SDS-polyacrylamide gel under non-reducing condition and stained with Coomassie blue.
Figure 3.
Confluent PAE/KDR cells were stimulated with 50ng/ml human VEGF165 for 10 min at 37°C. Cells were lysed and an IP was performed using a mouse anti-human KDR antibody (Cat# 101-M32). WB was performed with a mouse anti-human KDR antibody (Cat# 101-M34) and an anti-Phosphotyrosine antibody.
Figure 2.
Proliferation assay with primary human dermal lymphatic endothelial cells (HDLEC). The cells were stimulated using recombinant human VEGF165 and the WHO standard #02/286. Values are the means (±SD) of triplicate determinations and expressed as percentage of control.
Figure 4.
VEGF165-induced proliferation of HUVE cells. HUVECs were stimulated with increasing amounts of human VEGF165.

Reference

  1. Inter-manufacturer Comparison of Automated Immunoassays for the Measurement of Soluble FMS-like Tyrosine kinase-1 and Placental Growth Factor. Y. Kwun Yue Cheng et al., Pregnancy Hypertens. 2019 Jul;17:165-171.
  2. Pericyte-expressed Tie2 controls angiogenesis and vessel maturation. M. Teichert et al., Nat Commun. 2017; 8: 16106.
  3. MP0250, a VEGF and HGF neutralizing DARPin® molecule shows high anti-tumor efficacy in mouse xenograft and patient-derived tumor models. U. Fiedler et al., Oncotarget. 2017 Nov 17; 8(58): 98371–98383.
  4. Design and characterization of MP0250, a tri-specific anti-HGF/anti-VEGF DARPin® drug candidate. Binz HK et al., MAbs. 2017 Nov/Dec;9(8):1262-1269.
  5. Cryopreservation of primary human monocytes does not negatively affect their functionality or their ability to be labelled with radionuclides: basis for molecular imaging and cell therapy. E. Pardali et al., EJNMMI Res. 2016; 6: 77.
  6. The histone deacetylase inhibitor trichostatin a decreases lymphangiogenesis by inducing apoptosis and cell cycle arrest via p21-dependent pathways. I. Hrgovic et al., BMC Cancer. 2016; 16: 763.
  7. CD34 expression modulates tube-forming capacity and barrier properties of peripheral blood-derived endothelial colony-forming cells (ECFCs). D. Tasev et al., Angiogenesis. 2016; 19: 325–338.
  8. ADAM10 and ADAM17 have opposite roles during sprouting angiogenesis. Caolo V. et al., Angiogenesis. 2015 Jan;18(1):13-22.
  9. Endothelial Cells Derived from Non-malignant Tissues Are of Limited Value as Models for Brain Tumor Vasculature. Lohr J et al., Anticancer Res. 2015 May;35(5):2681-90.
  10. Long-Term Expansion in Platelet Lysate Increases Growth of Peripheral Blood-Derived Endothelial-Colony Forming Cells and Their Growth Factor-Induced Sprouting Capacity. D. Tasev et al., PLoS One. 2015; 10(6): e0129935.
  11. Improved Anchorage of Ti6Al4V Orthopaedic Bone Implants through Oligonucleotide Mediated Immobilization of BMP-2 in Osteoporotic Rats.  J. V. Wölfle et al., PLoS One. 2014; 9(1): e86151.
  12. Proteolytic Processing Regulates Placental Growth Factor Activities. D. C. Hoffmann et al., J Biol Chem. 2013 Jun 21; 288(25): 17976–17989.
  13. The Antiangiogenic 16K Prolactin Impairs Functional Tumor Neovascularization by Inhibiting Vessel Maturation. Ngoc-Quynh-Nhu Nguyen et al., PLoS One. 2011; 6(11): e27318.
  14. MicroRNA-21 Exhibits Antiangiogenic Function by Targeting RhoB Expression in Endothelial Cells. C. Sabatel et al., PLoS One. 2011; 6(2): e16979.
  15. Autocrine activity of soluble Flt-1 controls endothelial cell function and angiogenesis. S. Ahmad et al., Vasc Cell. 2011; 3: 15.
  16. Feed-forward Signaling by Membrane-bound Ligand Receptor Circuit: THE CASE OF NOTCH DELTA-LIKE 4 LIGAND IN ENDOTHELIAL CELLS. V. Caolo et al., J Biol Chem. 2010 Dec 24; 285(52): 40681–40689.
  17. Impaired Collateral Recruitment and Outward Remodeling in Experimental Diabetes. J. M. van Golde et al., Diabetes. 2008 Oct; 57(10): 2818–2823.
  18. Elevated placental soluble vascular endothelial growth factor receptor-1 inhibits angiogenesis in preeclampsia. S. Ahmad and A. Ahmed, Circ Res. 2004 Oct 29;95(9):884-91.
  19. Collagen type 1 retards tube formation by human microvascular endothelial cells in a fibrin matrix. Kroon, M.E. et al., Angiogenesis (2002) 5: 257.
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