Biotinylated Recombinant Human VEGF 165 Protein, CF

Biotinylated via amines
Catalog # Availability Size / Price Qty
BT10543-025
Biotinylated Recombinant Human VEGF 165 Protein Binding Activity
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Biotinylated Recombinant Human VEGF 165 Protein, CF Summary

Product Specifications

Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
When Recombinant Human VEGFR1/Flt-1 Fc Chimera Recombinant Human VEGF R1/Flt‑1 Fc Chimera (Catalog # 321-FL) is immobilized at 0.2 µg/mL (100 µL/well), Biotinylated Recombinant Human VEGF 165 (Catalog # BT10543) binds with an ED50 of 0.800-6.40 ng/mL.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human VEGF protein
Ala27-Arg191
Accession #
N-terminal Sequence
Analysis
Ala27
Structure / Form
Disulfide-linked homodimer, biotinylated via amines
Predicted Molecular Mass
19 kDa
SDS-PAGE
22-27 kDa, under reducing conditions

Product Datasheets

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BT10543

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

BT10543

Formulation Lyophilized from a 0.2 μm filtered solution in 4 mM HCl, pH 2.4 with Trehalose.
Reconstitution Reconstitute at 250 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.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Binding Activity Biotinylated Recombinant Human VEGF 165 Protein Binding Activity View Larger

When Recombinant Human VEGFR1/Flt-1 Fc Chimera (321-FL) is immobilized at 0.2 μg/mL (100 μL/well), Biotinylated Recombinant Human VEGF 165 (Catalog # BT10543) binds with an ED50 of 0.800-6.40 ng/mL.

SDS-PAGE Biotinylated Recombinant Human VEGF 165 Protein SDS-PAGE View Larger

2 μg/lane of Biotinylated Recombinant Human VEGF 165 Protein (Catalog # BT10543) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 22-27 kDa and 40-43 kDa, respectively.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: VEGF

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 (1-3). It is a member of the PDGF family that is characterized by the presence of eight conserved cysteine residues and a cystine knot structure (4). Humans express alternately spliced isoforms of 121, 145, 165, 183, 189, and 206 amino acids (aa) in length (4). VEGF165 appears to be the most abundant and potent isoform, followed by VEGF121 and VEGF189 (3, 4). Isoforms other than VEGF121 contain basic heparin-binding regions and are not freely diffusible (4). 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 VEGF, respectively. VEGF binds the type I transmembrane receptor tyrosine kinases VEGF R1 (also called Flt-1) and VEGF R2 (Flk-1/KDR) on endothelial cells (4). Although VEGF affinity is highest for binding to VEGF R1, VEGF R2 appears to be the primary mediator of VEGF angiogenic activity (3, 4). VEGF165 binds the semaphorin receptor, Neuropilin-1 and promotes complex formation with VEGF R2 (5). VEGF is required during embryogenesis to regulate the proliferation, migration, and survival of endothelial cells (3, 4). In adults, VEGF functions mainly in wound healing and the female reproductive cycle (3). Pathologically, it is involved in tumor angiogenesis and vascular leakage (6, 7). Circulating VEGF levels correlate with disease activity in autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and systemic lupus erythematosus (8). VEGF is induced by hypoxia and cytokines such as IL-1, IL-6, IL-8, oncostatin M and TNF-alpha (3, 4, 9). 

Due to its role in angiogenesis of blood vessels, tumor and stroma cells use VEGF to stimulate formation of blood vessels and the proliferation and survival of endothelial cells. Specific immunotherapies targeting the VEGF signaling pathway include the recombinant antibody against VEGF (Bevacizumab), antibodies targeting the main VEGF receptor (VEGFR2), and small molecule inhibitors against VEGF receptor tyrosine kinases (10). Immune checkpoint inhibitors are an important tool in cancer therapies as tumor cells can hijack immune checkpoint signals to evade detection by immune cells. In addition to stimulating the formation of tumor blood vessels, VEGF has immunosuppressive effects by acting on dendritic cells to block their antigen-presenting and T cell stimulatory functions. Targeting VEGF in combination with other immune checkpoint ligands or receptors may prove more effective in immunotherapy approaches to certain cancer types (11). Because of its role in the formation of blood vessels, VEGF is also an important factor in skeletal development where blood supply and vascularization are crucial. This has made VEGF an important molecule in regenerative studies for bone repair as sustained release of VEGF has been shown to improve the efficiency of bone regeneration (12). 

In differentiation protocols for stems cells, VEGF is a commonly added growth factor for the transformation of induced pluripotent stem cells into hematopoietic progenitor cells used to make Natural Killer cells (13,14). VEGF has also been used to transform intermediate mesoderm into kidney glomular podocytes or stem cell-derived liver spheres (15, 16). VEGF may also be used in assistance of stem cell transplantations by supporting angiogenesis at sites of stem cell transplants or as a honing tool for adipose-derived mesenchymal stem cells or bone marrow stem cells to migrate to (17,18).

References
  1. Leung, D.W. et al. (1989) Science 246:1306. 
  2. Keck, P.J. et al. (1989) Science 246:1309. 
  3. Byrne, A.M. et al. (2005) J. Cell. Mol. Med. 9:777. 
  4. Robinson, C.J. and S.E. Stringer (2001) J. Cell. Sci. 114:853.
  5. Pan, Q. et al. (2007) J. Biol. Chem. 282:24049.
  6. Weis, S.M. and D.A. Cheresh (2005) Nature 437:497.
  7. Thurston, G. (2002) J. Anat. 200:575.
  8. Carvalho, J.F. et al. (2007) J. Clin. Immunol. 27:246.
  9. Angelo, L.S. and R. Kurzrock (2007) Clin. Cancer Res. 13:2825.
  10. Apte, R.S. et al. (2019) Cell 176:1248.                                     
  11. Sangro, B. et al. (2021) Nature 18:525.                                        
  12. Hu, K. & Olsen, B.R. (2016) Bone 91:30.   
  13. Zhou, Y. et al. (2022) Cancers 14:2266.
  14. Li, Y. et al. (2018) Cell Stem Cell. 23:181.
  15. Musah, S. et al. (2018) Nat. Protoc. 13:1662.   
  16. Meseguer-Ripolles, J. et al. (2021) STAR Protoc. 2:100502.        
  17. Hutchings, G. et al. (2020) Int. J. Mol. Sci. 21:3790. 
  18. Zhang, W. et al. (2014) Eur. Cell. Mater. 27:1.
Long Name
Vascular Endothelial Growth Factor
Entrez Gene IDs
7422 (Human); 22339 (Mouse); 83785 (Rat); 281572 (Bovine); 403802 (Canine); 493845 (Feline); 30682 (Zebrafish)
Alternate Names
MVCD1; VAS; vascular endothelial growth factor A; Vascular permeability factor; Vasculotropin; VEGF; VEGFA; VEGF-A; VEGFMGC70609; VPF; VPFvascular endothelial growth factor

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