APPLICATION
Application Note
Vascular endothelial growth factor 165 is one kind of isoforms of Vascular endothelial growth factor A (VEGFA). This protein is a glycosylated mitogen that specifically acts on endothelial cells and has various effects, including mediating increased vascular permeability, inducing angiogenesis, vasculogenesis and endothelial cell growth, promoting cell migration, and inhibiting apoptosis. Besides, Vascular Endothelial Growth Factor Receptor 1 (VEGFR1) has been identified as an interactor of VEGF165, thus a binding ELISA assay was conducted to detect the interaction of recombinant human VEGF165 and recombinant human VEGFR1. Briefly, VEGF165 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to VEGFR1-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-VEGF165 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of VEGF165 and VEGFR1 was in a dose dependent manner.
Observed MW
19 kDa.
PROPERTIES
Form
Lyophilized powder
Buffer
Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
Preservative
ProClin 300
Storage
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
Region/Sequence
N-terminal His-Tag; Pro28~Arg191 (NP_001020537.2)
Expression System
E. coli
Purity
> 90%
Endotoxin
< 1 EU/μg
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET
Synonyms
vascular endothelial growth factor A , MVCD1 , VEGF , VPF
Background
This gene is a member of the PDGF/VEGF growth factor family. It encodes a heparin-binding protein, which exists as a disulfide-linked homodimer. This growth factor induces proliferation and migration of vascular endothelial cells, and is essential for both physiological and pathological angiogenesis. Disruption of this gene in mice resulted in abnormal embryonic blood vessel formation. This gene is upregulated in many known tumors and its expression is correlated with tumor stage and progression. Elevated levels of this protein are found in patients with POEMS syndrome, also known as Crow-Fukase syndrome. Allelic variants of this gene have been associated with microvascular complications of diabetes 1 (MVCD1) and atherosclerosis. Alternatively spliced transcript variants encoding different isoforms have been described. There is also evidence for alternative translation initiation from upstream non-AUG (CUG) codons resulting in additional isoforms. A recent study showed that a C-terminally extended isoform is produced by use of an alternative in-frame translation termination codon via a stop codon readthrough mechanism, and that this isoform is antiangiogenic. Expression of some isoforms derived from the AUG start codon is regulated by a small upstream open reading frame, which is located within an internal ribosome entry site. [provided by RefSeq, Nov 2015]
Database
Research Area
DATA IMAGES
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GTX00110-pro Functional Assay Image
Functional ELISA analysis of GTX00110-pro Human VEGF165 protein which can bind immobilized VEGFR1 protein.
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GTX00110-pro Image
SDS-PAGE analysis of GTX00110-pro Human VEGF165 protein.
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GTX00110-pro Image
WB analysis of GTX00110-pro Human VEGF165 protein.
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REFERENCE
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REVIEW
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