Recombinant Human VEGF 165 GMP Protein, CF GMP

Name: Recombinant Human VEGF 165 GMP Protein, CF
Brand: R&D Systems
SKU: BT-VEGF-GMP

Catalog # BT-VEGF-GMP | R&D Systems, Inc. a Bio-Techne Brand

Product Datasheet / COA / SDS

Animal-Free

Setting the standard in quality research reagents

Key Product Details

VEGF Manufactured in Bio-Techne's new GMP facility
Lot-to-lot consistency
Stringent guidelines for patient safety
Scalability necessary to support successful therapeutics
Learn more about manufacturing in our new GMP facility
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Source:	E. coli
Accession #:	NP_001165097.1
Structure / Form:	Disulfide-linked homodimer
Applications:	Bioactivity

View Full Technical Details for Recombinant Human VEGF 165 GMP Protein, CF

Technical Data
Citations / Reviews

Product Specifications for Recombinant Human VEGF 165 GMP Protein, CF

Source	E. coli-derived human VEGF protein Ala27 - Arg191 Produced using non-animal reagents in an animal-free laboratory. Manufactured and tested under cGMP guidelines.
Purity	>97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. The molecular weight by TCEP reduced mass spectrometry is 19282 ± 10 Da and 19080 Da ± 10 Da, and is free of unexpected mass peaks
Endotoxin level	<0.01 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Met-Ala27-Pro-Met-Ala-Glu-Gly-Gly-Gly-Gln Pro28-Met-Ala-Glu-Gly-Gly-Gly-Gln-Asn-His
Predicted Molecular Mass	19.2 kDa (monomer)
SDS-PAGE	19.2 kDa (monomer)
Activity	Measured in a cell proliferation assay using HUVEC human umbilical vein endothelial cells. Conn, G. et al. (1990) Proc. Natl. Acad. Sci. USA 87:1323. The ED₅₀ for this effect is 1.50-12.0 ng/mL. The specific activity of recombinant human VEGF₁₆₅ is > 8.0 x 10⁵ units/mg, which is calibrated against the human VEGF₁₆₅ WHO standard (NIBSC code: 02/286).

Scientific Data Examples for Recombinant Human VEGF 165 GMP Protein, CF

	Recombinant Human VEGF 165 GMP Protein Bioactivity. GMP-grade Recombinant Human VEGF 165 (Catalog # BT-VEGF-GMP) as measured in a cell proliferation assay using HUVEC human umbilical vein endothelial cells. The ED₅₀ for this effect is 1.50-12.0 ng/mL. Three independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of GMP VEGF 165.
	Equivalent Bioactivity of GMP and Animal-Free grades of Recombinant Human VEGF 165. Equivalent bioactivity of GMP (Catalog # BT-VEGF-GMP) and Animal-Free (BT-VEGF-AFL) grades of Recombinant Human VEGF 165 as measured in a cell proliferation assay using HUVEC human umbilical vein endothelial cells (orange and green, respectively).
	Recombinant Human VEGF 165 GMP Protein SDS-PAGE 2 μg/lane of Recombinant Human VEGF 165 GMP Protein (Catalog # BT-VEGF-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 19-21 kDa.
	Recombinant Human VEGF 165 GMP Protein Purity Analysis by Size Separation. Three independent lots of Recombinant Human VEGF 165 GMP Protein (Catalog # BT-VEGF-GMP) were analyzed by Maurice CE-SDS PLUS (IS is an Internal Standard). A gel-like representation of the purity analysis data (inset) can be obtained from the Lane View feature in Compass software for iCE. Profiles from the three runs were superimposed, showing excellent manufacturing consistency.
	Recombinant Human VEGF 165 GMP Protein Purity Analysis by Charge Variation. Three independent lots of Recombinant Human VEGF 165 GMP Protein (Catalog # BT-VEGF-GMP) were analyzed by Maurice icIEF using native fluorescence detection (Mkr 5.85 and 9.99 are pI Markers). Profiles from the three runs were superimposed, showing excellent manufacturing consistency.

Formulation, Preparation and Storage

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.

BT-VEGF-GMP

Formulation	Lyophilized from a 0.2 μm filtered solution in Sodium Acetate.
Reconstitution	Reconstitute at 500 μg/mL in sterile deionized water.
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. A minimum of 12 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, ≤ -20 °C under sterile conditions after reconstitution.

Reconstitution Calculator

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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 glomerular 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).

Item	Value
References	Leung, D.W. et al. (1989) Science 246:1306. Keck, P.J. et al. (1989) Science 246:1309. Byrne, A.M. et al. (2005) J. Cell. Mol. Med. 9:777. Robinson, C.J. and S.E. Stringer (2001) J. Cell. Sci. 114:853. Pan, Q. et al. (2007) J. Biol. Chem. 282:24049. Weis, S.M. and D.A. Cheresh (2005) Nature 437:497. Thurston, G. (2002) J. Anat. 200:575. Carvalho, J.F. et al. (2007) J. Clin. Immunol. 27:246. Angelo, L.S. and R. Kurzrock (2007) Clin. Cancer Res. 13:2825. Apte, R.S. et al. (2019) Cell 176:1248. Sangro, B. et al. (2021) Nature 18:525. Hu, K. & Olsen, B.R. (2016) Bone 91:30. Zhou, Y. et al. (2022) Cancers 14:2266. Li, Y. et al. (2018) Cell Stem Cell. 23:181. Musah, S. et al. (2018) Nat. Protoc. 13:1662. Meseguer-Ripolles, J. et al. (2021) STAR Protoc. 2:100502. Hutchings, G. et al. (2020) Int. J. Mol. Sci. 21:3790. 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, VEGF, VEGF-A, VEGFA, VEGFMGC70609, VPF, VPFvascular endothelial growth factor, Vascular permeability factor, Vasculotropin, vascular endothelial growth factor A

Manufacturing Specifications

GMP Proteins
R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Eu. Ph. 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.

R&D Systems' quality focus includes:

Manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
Documented processes and QA control of documentation and process changes
Personnel training programs
Raw material testing and vendor qualification/monitoring
Fully validated equipment, processes and test methods
Equipment calibration schedules using a computerized calibration program
Facility maintenance, safety programs and pest control
Material review process for variances
Monitoring of stability over product shelf-life

R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their research. The Certificate of Analysis provided contains the following lot specific information:

N-terminal amino acid analysis, SDS-PAGE analysis, and endotoxin level (as determined by LAL assay) performed on each bulk QC lot, not on individual bottlings of each QC lot
Post-bottling lot-specific bioassay results (compliance with an established range) and results of microbial testing according to USP
Host Cell Protein testing performed by ELISA
Mycoplasma testing by ribosomal RNA hybridization assay

Additional testing and documentation requested by the customer can be arranged at an additional cost.

Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis, Minnesota USA.

R&D Systems sells GMP grade products for preclinical or clinical ex vivo use. They are not for in vivo use. Please read the following End User Terms prior to using this product.

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.

Production

All molecular biology procedures use animal-free media and dedicated labware.
Dedicated fermentors are utilized in committed animal-free areas.

Purification

Protein purification columns are animal-free.
Bulk proteins are filtered using animal-free filters.
Purified proteins are stored in animal-free containers in a dedicated cold storage room.

Quality Assurance

Low Endotoxin Level.
No impairment of biological activity.
High quality product obtained under stringent conditions.

Please read our complete Animal-Free Statement.

Product Specific Notices for Recombinant Human VEGF 165 GMP Protein, CF

END USER TERMS OF USE OF PRODUCT

The following terms are offered to you upon your acceptance of these End User Terms of Use of Product. By using this product, you indicate your acknowledgment and agreement to these End User Terms of Use of Product. If you do not agree to be bound by and comply with all of the provisions of these End User Terms of Use of Product, you should contact your supplier of the product and make arrangements to return the product.

We suggest you print and retain a copy of these End User Terms of Use of Product for your records.

The End User is aware that R&D Systems, Inc. sells GMP products for preclinical or clinical ex vivo use and not for in vivo use. The End User further agrees, as a condition of the sale of R&D Systems' GMP products that: a) the End User will not use this GMP Product in any procedure wherein the product may be directly or indirectly administered to humans, unless the End User has obtained, or prior to their use will have obtained, an Investigational New Drug (IND) exemption from the FDA and will use the product only in accordance with the protocols of such IND and of the Institutional Review Board overseeing the proposed research, or b) the End User will use the products outside of the United States in accordance with the protocols of research approved by the Institutional Review Board or authorized ethics committee and regulatory agencies to which the End User is subject to in their territory.

R&D Systems, Inc. has the right, at its sole discretion, to modify, add or remove any terms or conditions of these End User Terms of Use without notice or liability to you. Any changes to these End User Terms of Use are effective immediately following the printing of such changes on this product insert. The most recent version of these End User Terms of Use of Product may be found at: RnDSystems.com/Legal.

You agree to review these End User Terms of Use of Product to ensure any subsequent use by you of R&D Systems' GMP Products following changes to these End User Terms of Use of Product constitutes your acceptance of all such changes.

TERMS AND CONDITIONS

The following limitation applies to R&D Systems' warranty and liability for damages: All products are warranted to meet R&D Systems' published specifications when used under normal laboratory conditions.

R&D SYSTEMS DOES NOT MAKE ANY OTHER WARRANTY OR REPRESENTATION WHATSOEVER, WHETHER EXPRESS OR IMPLIED, WITH RESPECT TO ITS PRODUCTS. IN PARTICULAR, R&D SYSTEMS DOES NOT MAKE ANY WARRANTY OF SUITABILITY, NONINFRINGEMENT, MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE.

NOTWITHSTANDING ANY OTHER PROVISIONS OF THESE TERMS AND/OR ANY OTHER AGREEMENT BETWEEN R&D SYSTEMS AND PURCHASER FOR THE PURCHASE OF THE PRODUCTS, R&D SYSTEMS' TOTAL LIABILITY TO PURCHASER ARISING FROM OR IN RELATION TO THESE TERMS, AN AGREEMENT BETWEEN THE PARTIES OR THE PRODUCTS, WHETHER ARISING IN CONTRACT, TORT OR OTHERWISE SHALL BE LIMITED TO THE TOTAL AMOUNT PAID BY PURCHASER TO R&D SYSTEMS FOR THE APPLICABLE PRODUCTS. IN NO EVENT WILL R&D SYSTEMS BE LIABLE FOR THE COST OF PROCUREMENT OF SUBSTITUTE GOODS.

Full details of R&D Systems' Terms and Conditions of Sale can be found online at: RnDSystems.com/Legal.

Citations for Recombinant Human VEGF 165 GMP Protein, CF

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