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Recombinant human VEGF165 protein (Qk048)

Recombinant human vascular endothelial growth factor 165 (also known as VEGF165/ VEGF-165/ VEGF-A/ VEGF165) protein is widely used in culturing primary endothelial cells (ECs), such as human umbilical vein endothelial cells (HUVEC), under serum-free conditions for blood vessel developmental studies. VEGF165 is commonly used with human-induced pluripotent stem cells (hiPSC) or embryonic stem cells (ESC)-derived EC for developing human vascular tissue models for disease mechanism studies. VEGF165 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.

A highly bioactive 38 kDa homodimer, which is animal-derived component-free (ADCF) and carrier protein-free (CF).

Orders are typically shipped same or next day (except Friday).
Easy world-wide ordering, direct or through our distributors.

1mg may be despatched as 2 x 500µg

Buy online with secure credit card or purchase order or email orders@qkine.com
Bulk and stock reservation available

Summary

  • High purity human VEGF165 (Uniprot: P15692)

  • 38 kDa dimer, 19 kDa monomer

  • Expressed in E. coli
  • 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

Bioactivity

Bioactivity graph showing the EC50 of 0.55 ng/ml (14.4 pM) for Qkine recombinant human VEGF 165

The bioactivity of VEGF165 is measured using a luciferase reporter cell line which stably expresses the KDR (VEGFR-2) receptor. Cells were incubated with different concentrations of VEGF165 for 6 hours before assaying for luciferase production.

EC50=0.55 ng/ml (14.4 pM), data from Qk048 lot #104393, n=3.

Purity

SDS-PAGE gel showing high purity reduced and non-reduced forms of VEGF 165

Human VEGF165 (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 VEGF165 has higher bioactivity than an alternative supplier.

The bioactivity of Qkine and an alternative supplier of VEGF165 were compared directly using a luciferase reporter cell line which stably expresses the KDR (VEGFR-2) receptor. Cells were incubated with different concentrations of VEGF165 for 6 hours before assaying for luciferase production.

EC50=0.55 ng/ml (14.4 pM) for Qk048 VEGF165, data from Qk048 lot #104393, n=3

EC50=1.44 ng/ml (37.6 pM) for Supplier 1 VEGF165, , n=3

Protein background

Vascular endothelial growth factor (VEGF) is a member of the platelet-derived growth factor (PDGF) family and signals through the type I transmembrane receptor tyrosine kinases VEGFR1 (also called Flt-1) and VEGFR2 (Flk-1/KDR). 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 [1].
VEGF165 is a core regulator of angiogenesis and vasculogenesis. It is responsible for the survival, proliferation, migration, and specialization of endothelial cells, thus called an endothelial cell surviving factor [2]. VEGF 165 has multiple roles in the CNS, such as stimulating neural stem cell (NSC) proliferation and promoting neurogenesis [3]. VEGF enhances the migration of oligodendrocyte precursor cells, increases migration and proliferation of Schwann cells, stimulates expression of trophic factors by astrocytes, and triggers proliferation, survival and migration of astrocytes and microglia [4]. It is a critical factor in generating human pluripotent stem cell- (iPSC- or ESC-) derived vascularized brain organoids [5]. Immunocytochemical staining with CD31 expression marker is used to indicate the presence of endothelial cells, thereby suggesting successful endothelial cell differentiation [5]. VEGF also 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 [6].
In humans, VEGF is produced as multiple alternately spliced isoforms. Nine major VEGF‐A isoforms have been identified in humans: VEGF121, VEGF145, VEGF148, VEGF162, VEGF165, VEGF165b (an anti-angiogenic isoform that binds to VEGFR2 with similar affinity to VEGF165 but does not activate it), VEGF183, VEGF189, and VEGF206.
VEGF165 is the most abundantly expressed isoform [7], also known as VEGF-A 165a. Qkine recombinant human VEGF165 protein is a highly pure and potent 38 kDa homodimer, disulfide-linked consisting of two 165 amino acid polypeptide chains. It is an animal-derived component-free (ADCF) and carrier protein-free (CF) isoform of VEGF for high efficiency.

1. Sulpice, E. et al. Neuropilin-1 and neuropilin-2 act as coreceptors, potentiating proangiogenic activity. Blood 111, 2036–2045 (2008).
2. Honnegowda, T. M. et al. Role of angiogenesis and angiogenic factors in acute and chronic wound healing. Plastic and Aesthetic Research 2, 243–249 (2015).
3. Storkebaum E et al. VEGF: once regarded as a specific angiogenic factor, now implicated in neuroprotection. Bioessays 26(9):943–54 (2004).
4. Lange, C., Storkebaum, E., de Almodóvar, C. et al. Vascular endothelial growth factor: a neurovascular target in neurological diseases. Nat Rev Neurol 12, 439–454 (2016).
5. Pham, Missy T., et al. Generation of human vascularized brain organoids. Neuroreport 29.7: 588 (2018).
6. 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).
7. Bhisitkul, R. B. Vascular endothelial growth factor biology: clinical implications for ocular treatments. Br J Ophthalmol 90, 1542–1547 (2006).

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

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