Recombinant Human VEGF 165 Biotinylated Protein, CF
Recombinant Human VEGF 165 Biotinylated Protein, CF Summary
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|Formulation||Lyophilized from a 0.2 μm filtered solution in Acetonitrile and TFA.|
|Reconstitution||Reconstitute at 100 μg/mL in 4 mM HCl.|
|Shipping||The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
Both Biotinylated Recombinant Human VEGF 165 (Catalog # BT293/CF) and unlabeled Recombinant Human VEGF 165 (Catalog # 293-VE) stimulate HUVEC human umbilical vein endothelial cell proliferation. The ED50 for this effect is 1-6 ng/mL. The similarity in activity highlights that the biotinylayed protein is fully functional.
Vascular endothelial growth factor (VEGF or VEGF-A), also known as vascular permeability factor (VPF), is a potent mediator of both angiogenesis and vasculogenesis in the fetus and adult (1, 2). It is a member of the PDGF family that is characterized by the presence of eight conserved cysteine residues and a cystine knot structure (3). Humans express two sets of alternatively spliced isoforms of 121, 145, 165, 183, 189, and 206 amino acids (aa) in length (3, 4). Isoforms other than VEGF121 contain basic heparin-binding regions and are not freely diffusible (3, 4). VEGF165 appears to be the most abundant and potent of the angiogenic isoform set, followed by VEGF121 and VEGF189 (3, 5). The anti-angiogenic or “b” set of isoforms is differentially spliced to contain five alternative amino acids at the C-terminus, and are the more highly expressed isoforms in normal adult tissue (6). VEGF165b, like VEGF121 but unlike most angiogenic isoforms, does not bind heparins and is therefore diffusible (3). Human VEGF165 shares 88% aa sequence identity with corresponding regions of mouse and rat, 96% with porcine, 95% with canine, and 93% with feline, equine and bovine VEGF165, respectively. In addition to alternatively spliced VEGF isoforms, multiple fragments of VEGF can be generated by extracellular proteolysis (4). VEGFs bind the type I transmembrane receptor tyrosine kinases VEGF R1 (also called Flt-1) and VEGF R2 (Flk-1/KDR) on endothelial cells (3). Although VEGF affinity is highest for binding to VEGF R1, VEGF R2 appears to be the primary mediator of VEGF angiogenic activity (3, 5). VEGF165 binds the semaphorin receptor, Neuropilin-1 and promotes complex formation with VEGF R2 (7). VEGF is required during embryogenesis to regulate the proliferation, migration, and survival of endothelial cells (3, 5). In adults, VEGF functions mainly in wound healing and the female reproductive cycle (5). Pathologically, it is involved in tumor development and tumor vascular leakage (8). Circulating VEGF levels correlate with disease activity in autoimmune diseases such as rheumatoid arthritis, multiple sclerosis, and systemic lupus erythematosus (9). VEGF is induced by hypoxia and cytokines such as IL-1, IL-6, IL-8, Oncostatin M, and TNF-alpha (5, 10).
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- Robinson, C.J. and S.E. Stringer (2001) J. Cell. Sci. 114:853.
- Vempati, P. et al. (2014) Cytokine Growth Factor Rev. 25:1.
- Byrne, A.M. et al. (2005) J. Cell. Mol. Med. 9:777.
- Nowak, D.G. et al. (2008) J. Cell Sci. 121:3487.
- Pan, Q. et al. (2007) J. Biol. Chem. 282:24049.
- Goel, H.L. and A.M. Mercurio (2013) Nat. Rev. Cancer 13:871.
- Carvalho, J.F. et al. (2007) J. Clin. Immunol. 27:246.
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Citations for Recombinant Human VEGF 165 Biotinylated Protein, CF
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.
Citations: Showing 1 - 3
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Islet vascularization is regulated by primary endothelial cilia via VEGF-A-dependent signaling
Authors: Y Xiong, MJ Scerbo, A Seelig, F Volta, N O'Brien, A Dicker, D Padula, H Lickert, JM Gerdes, PO Berggren
Sample Types: Whole Cells
Applications: Cell Culture
Disrupting VEGF-VEGFR1 Interaction: De Novo Designed Linear Helical Peptides to Mimic the VEGF13-25 Fragment
Authors: B Balsera, MÁ Bonache, M Reille-Ser, N Gagey-Eils, M Vidal, R González-M, MJ Pérez de V
Sample Types: Recombinant Protein
Applications: Binding Assay
DSGOST inhibits tumor growth by blocking VEGF/VEGFR2-activated angiogenesis
Sample Types: Protein
The vial is supposed to contain lyophilized protein but it appears to be empty. Is there anything in it?
Pellets can be dislodged during shipping and become disbursed on the vial wall and in the cap. Centrifuge or tap the vial on the benchtop to return this material to the vial bottom. If this does not reveal a pellet, closely inspect the cone of the vial. Some pellets appear as only a tiny amount of material or as a transparent film due to the original buffer formulation. This is a normal appearance for many proteins. For example, if the product is originally lyophilized from a solvent such as acetonitrile or ethanol, and supplied carrier-free, you may not be able to detect the pellet with the naked eye. This does not mean the vial is empty. Reconstitute the vial as directed. After reconstitution, protein concentration can be tested with a spectrophotometer.
What is the recommended method for reconstitution of a lyophilized protein or antibody?
Unless more specific directions are on the Certificate of Analysis provided with the product, we suggest the following procedure to ensure optimal recovery: 1. Allow the vial and reconstitution buffer to equilibrate to room temperature. 2. Briefly centrifuge the vial to ensure that all lyophiliate is collected at the bottom of the vial. 3. Add the amount of buffer required to achieve the concentration recommended on the product insert. 4. Allow the vial to reconstitute for 15-30 minutes at room temperature with gentle agitation, like on a rocker platform or rotating by hand. Avoid vigorous shaking that can cause foaming and protein denaturation. 5. Aliquot into volumes greater than 20 μL and store as indicated on the product insert. If the vial exhibits flakes or particulates, mix the product for a couple of hours at room temperature and then at 4oC overnight. Contact Technical Service if product does not go into solution.
Are R&D Systems recombinant proteins and antibodies sterile?
Although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is needed for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.
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