Recombinant human VEGF 165 protein (Qk048)

Recombinant human vascular endothelial growth factor 165 (VEGF165/ VEGF-165/ VEGF165) protein is widely used in culturing primary endothelial cells, such as human umbilical vein endothelial cells (HUVEC).

VEGF 165 is commonly used with human-induced pluripotent stem cells or embryonic stem cells-derived endothelial cells for developing human vascular tissue models. It has many applications including its use in neural research involving oligodendrocyte precursor cells, Schwann cells, astrocytes, and microglia. It plays a role in bone formation, regulates mesenchymal stem cell differentiation, and serves as a survival factor for chondrocytes, hematopoietic stem cells, and tumor cells.

Qkine recombinant human VEGF 165 protein is a highly pure and potent 38 kDa homodimer, disulfide-linked consisting of two 165 amino acid polypeptide chains. It is an animal origin-free (AOF) and carrier protein-free (CF).

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Summary

  • High purity human VEGF 165 (Uniprot: P15692)

  • >98%, by SDS-PAGE quantitative densitometry

  • 38.3 kDa (dimer), 19 kDa monomer

  • Expressed in E. coli
  • Animal origin-free (AOF) and carrier protein-free

  • Manufactured in Cambridge, UK

  • Lyophilized from acetonitrile/TFA
  • Resuspend in water at >100 µg/ml, prepare single use aliquots, add carrier protein if desired and store frozen at -20 oC (short-term) or -80 oC (long-term)

Featured applications

  • Angiogenic cell research

  • Endothelial cell differentiation
  • Vasculature in organoids
  • Neural stem cells research
  • Mesenchymal stem cells research
  • hPSC-derived mesoderm differentiation

Vascular Endothelial Growth Factor, VPF, Folliculostellate cell-derived growth factor, Glioma-derived endothelial cell mitogen, MGC70609, MVCD-1, Vascular endothelial growth factor 2, Vascular endothelial growth factor A, Vascular permeability factor, VEGF­A, VPF, MVCD1, VAS, VEGFMGC70609, VEGF, VEGFA

human

species similarity:
mouse – 88%
rat – 88%
porcine – 97%
bovine – 94%

Bioactivity

Qkine recombinant human VEGF 165 - Bioactivity graph

The bioactivity of Qk048 is measured using a luciferase reporter cell line which stably expresses the KDR (VEGFR-2) receptor. Cells were incubated with different concentrations of VEGF 165 for 6 hours before assaying for luciferase production. EC50=0.55 ng/ml (14.4 pM), data from Qk048 lot #104393, n=3.

Purity

VEGF 165 SDS-PAGE showing high purity

Human VEGF 165 (Qk048) migrates as a dimer at 38 kDa in non-reducing (NR) conditions and as a monomer at 19 kDa upon reduction (R). No contaminating bands are visible. Purified recombinant protein (3 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptoethanol, R) and non-reduced (-β-mercaptoethanol, NR) conditions and stained with Coomassie Brilliant Blue R-250. Data from Qk048 lot #104393.

Further quality assays

  • Mass spectrometry: single species with expected mass

  • Endotoxin: <0.005 EU/μg protein (below level of detection)

  • Recovery from stock vial:  >95%

We are a company founded and run by scientists to provide a service and support innovation in stem cell biology and regenerative medicine.  All our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.

Graph showing a higher bioactivity (lower EC50) of Qkine recombinant human VEGF 165 protein in comparison to an alternative supplier

Comparison with other suppliers

Qkine VEGF 165 has higher bioactivity than an alternative supplier

The bioactivity of Qkine and an alternative supplier were compared directly using a luciferase reporter cell line which stably expresses the KDR (VEGFR-2) receptor. Cells were incubated with different concentrations of VEGF 165 for 6 hours before assaying for luciferase production. EC50=0.55 ng/ml (14.4 pM) for Qk048, data from Qk048 lot #104393, n=3. EC50=1.44 ng/ml (37.6 pM) for Supplier 1 VEGF 165, , n=3.

Protein background

Vascular endothelial growth factor (VEGF) is a member of the platelet-derived growth factor (PDGF) family and a core regulator of angiogenesis and vascular permeability [1,2]. It is responsible for the survival, proliferation, migration, and specialization of endothelial cells, thus called an endothelial cell surviving factor [2,3].

In humans, VEGF is produced as multiple alternately spliced isoforms indicating the number of amino acids in length: VEGF121, VEGF145, VEGF148, VEGF162, VEGF165a, VEGF165b, VEGF183, VEGF189, and VEGF206. VEGF165 (or VEGF165a) is the most abundantly expressed isoform composed of 165 amino acids [1].

VEGF165 signals through the type I transmembrane receptor tyrosine kinases VEGFR1 (also called Flt-1) and VEGFR2 (Flk-1/KDR) [2]. Some VEGF family members (including VEGFA 165) also bind to the co-receptors neuropilin 1 (NRP1) and neuropilin 2 (NRP2), which can stimulate VEGFR2 activation. It is characterized by the presence of eight conserved cysteine residues forming a receptor-binding cystine-knot structure [4,5].

VEGF is widely used in culturing primary endothelial cells, such as human umbilical vein endothelial cells, under serum-free conditions for blood vessel developmental studies. VEGF165 is commonly used with human-induced pluripotent stem cells or embryonic stem cells-derived endothelial cells for developing human vascular tissue models for disease mechanism studies. Immunocytochemical staining with CD31 expression marker is used to indicate the presence of endothelial cells, thereby suggesting successful endothelial cell differentiation [6,7]. Endothelial cells can also be derived from hair follicle stem cells [8].

In addition to its angiogenic role, it is also involved in promoting neurogenesis and stimulating neural stem cell proliferation [9,10]. It can promote the proliferation, survival or migration of other glial cells such as oligodendrocyte precursor cells, Schwann cells, and can stimulate the expression of trophic factors by astrocytes [11]. Moreover, it is a critical factor in generating human pluripotent stem cell-derived vascularized brain organoids [6].

Finally, VEGF also plays a role in bone formation and regulates mesenchymal stem cell differentiation of pluripotent stem cells and bone marrow stem cells [12,13]. Therefore, it serves as a survival factor for chondrocytes, hematopoietic stem cells, and tumor cells [12,14].

The strong implications of VEGF have led to preclinical studies on the potential of VEGF administration in neurodegenerative and ischemic diseases [1,2,11]. The inhibition of VEGF has been approved for the treatment of neovascular ocular disease to prevent the blood brain barrier breakdown or excessive angiogenesis11. It is also a target in anti-angiogenic strategies in cancer due to its contribution in tumor angiogenesis [2,14].

[1] Bhisitkul, R. B. Vascular endothelial growth factor biology: clinical implications for ocular treatments. Br. J. Ophthalmol. 90, 1542–1547 (2006). doi: 10.1136/bjo.2006.098426

[2] Shibuya, M. Vascular Endothelial Growth Factor (VEGF) and Its Receptor (VEGFR) Signaling in Angiogenesis. Genes Cancer 2, 1097–1105 (2011). doi: 10.1177/1947601911423031

[3] Honnegowda, T. M., Kumar, P. & Rao, P. Role of angiogenesis and angiogenic factors in acute and chronic wound healing. Plast. Aesthetic Res. 2, null (2015). doi: 10.4103/2347-9264.165438

[4] Ferrara, N., Gerber, H.-P. & LeCouter, J. The biology of VEGF and its receptors. Nat. Med. 9, 669–676 (2003). doi.org/10.1038/nm0603-669

[5] Sulpice, E. et al. Neuropilin-1 and neuropilin-2 act as coreceptors, potentiating proangiogenic activity. Blood 111, 2036–2045 (2008). doi: 10.1182/blood-2007-04-084269

[6] Pham, M. T. et al. Generation of human vascularized brain organoids. Neuroreport 29, 588–593 (2018). doi: 10.1097/WNR.0000000000001014

[7] Atchison, L. et al. iPSC-Derived Endothelial Cells Affect Vascular Function in a Tissue-Engineered Blood Vessel Model of Hutchinson-Gilford Progeria Syndrome. Stem Cell Rep. 14, 325–337 (2020). doi: 10.1016/j.stemcr.2020.01.005

[8] Quan, R. et al. VEGF165 induces differentiation of hair follicle stem cells into endothelial cells and plays a role in in vivo angiogenesis. J. Cell. Mol. Med. 21, 1593–1604 (2017). doi: 10.1111/jcmm.13089

[9] Schänzer, A. et al. Direct Stimulation of Adult Neural Stem Cells In Vitro and Neurogenesis In Vivo by Vascular Endothelial Growth Factor. Brain Pathol. 14, 237–248 (2004). doi: 10.1111/j.1750-3639.2004.tb00060.x

[10] Storkebaum, E., Lambrechts, D. & Carmeliet, P. VEGF: once regarded as a specific angiogenic factor, now implicated in neuroprotection. BioEssays 26, 943–954 (2004). doi: 10.1002/bies.20092

[11] Lange, C., Storkebaum, E., de Almodóvar, C. R., Dewerchin, M. & Carmeliet, P. Vascular endothelial growth factor: a neurovascular target in neurological diseases. Nat. Rev. Neurol. 12, 439–454 (2016). doi: 10.1038/nrneurol.2016.88

[12] Berendsen, A. D. & Olsen, B. R. How Vascular Endothelial Growth Factor-A (VEGF) Regulates Differentiation of Mesenchymal Stem Cells. J. Histochem. Cytochem. 62, 103–108 (2014). doi: 10.1369/0022155413516347

[13] Lin, Z. et al. Effects of BMP2 and VEGF165 on the osteogenic differentiation of rat bone marrow‑derived mesenchymal stem cells. Exp. Ther. Med. 7, 625–629 (2014). doi: 10.3892/etm.2013.1464

[14] Duffy, A. M., Bouchier-Hayes, D. J. & Harmey, J. H. Vascular Endothelial Growth Factor (VEGF) and Its Role in Non-Endothelial Cells: Autocrine Signalling by VEGF. in Madame Curie Bioscience Database [Internet] (Landes Bioscience, 2013). https://www.ncbi.nlm.nih.gov/books/NBK6482/

Publications using recombinant human VEGF 165 protein (Qk048)

STAT3 signalling enhances tissue expansion during postimplantation mouse development
bioRxiv preprint (2024) by Azami, T. et al.

De novo design of high-affinity protein binders with AlphaProteo.
arXiv on 12 September 2024 by Zambaldi, V., La, D., Chu, A. E. et al.

FAQ

Vascular Endothelial Growth Factor 165 is a specific isoform of the vascular endothelial growth factor family. It stimulates vascular permeability plays a crucial role in various embryonic development, wound healing, and tumor angiogenesis.

VEGF 165 stimulates angiogenesis by stimulating the proliferation of endothelial cells. It is also involved in the regulation and differentiation of pluripotent stem cells into endothelial cells, neural stem cells, and hematopoietic stem cells.

Angiogenesis is the process by which new blood vessels are formed. It is a critical part of embryonic development, growth, and wound healing. It is regulated by pro-angiogenic factors such as VEGF acting on endothelial cells, cells lining the interior of blood vessels.

VEGF 165 and VEGF 121 are two different alternatively spliced isoforms of the vascular endothelial growth factor (VEGF) protein. VEGF 165 and VEGF 121 are composed of 165 and 121 amino acids respectively.

VEGF 165 binds to the VEGF receptor tyrosine kinases, known as VEGFR-1 (Flt-1) and VEGFR-2 (KDR/Flk-1) on endothelial cells.

Our products are for research use only and not for diagnostic or therapeutic use.  Products are not for resale.

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