01-185
VEGF Protein, Human recombinant
The human recombinant VEGF Protein is available in a 10 µg format.
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About This Item
UNSPSC Code:
12352202
eCl@ss:
32160405
NACRES:
NA.75
Recommended Products
biological source
human
Quality Level
recombinant
expressed in baculovirus infected Sf9 cells
form
liquid
manufacturer/tradename
Upstate®
technique(s)
cell culture | mammalian: suitable
input
sample type neural stem cell(s)
sample type hematopoietic stem cell(s)
sample type mesenchymal stem cell(s)
NCBI accession no.
UniProt accession no.
shipped in
dry ice
Gene Information
human ... VEGFA(7422)
General description
Product Source: Recombinant human DNA expressed in baculovirus infected Sf9 cells
Application
VEGF Proliferation Assay
1. Prior to use in this assay, maintain HUVE cells in F-12K medium supplemented with 10% (v/v) FBS, 1% penicillin-streptomycin-glutamine, 0.1 mg/ml heparin and 0.05 mg/ml endothelial cell growth supplement (ECGS).
2. For assay of VEGF, subculture HUVE cells into a 96-well plate at a density of 4,000 cells per well in the growth medium over night.
3. Add VEGF to a final concentration range of 0.2, 0.4, 0.8, 1.6, 3.2, 6.2, 12.5, 25 and 50ng/ml. Add VEGF diluent to one set of wells as the negative control.
4. Four days later, add 20ul of Promega Substrate Cell Titer 96 Aqueous One Solution Reagent to each well.
5. Incubate at 37C for 2 hrs and read at OD 490nm.
1. Prior to use in this assay, maintain HUVE cells in F-12K medium supplemented with 10% (v/v) FBS, 1% penicillin-streptomycin-glutamine, 0.1 mg/ml heparin and 0.05 mg/ml endothelial cell growth supplement (ECGS).
2. For assay of VEGF, subculture HUVE cells into a 96-well plate at a density of 4,000 cells per well in the growth medium over night.
3. Add VEGF to a final concentration range of 0.2, 0.4, 0.8, 1.6, 3.2, 6.2, 12.5, 25 and 50ng/ml. Add VEGF diluent to one set of wells as the negative control.
4. Four days later, add 20ul of Promega Substrate Cell Titer 96 Aqueous One Solution Reagent to each well.
5. Incubate at 37C for 2 hrs and read at OD 490nm.
Quality
Routinely evaluated in a 5-day growth assay using human umbilical vein endothelial cells
Cell Proliferation Assay: The biological activity of this lot of VEGF was determined by its mitogenic activity on human umbilical vein endothelial cells (HUVEC) with an ED50 of about 1.0 ng/mL. Results may vary depending on the cell line used. VEGF will also stimulate endothelial cells from other species.
Cell Proliferation Assay: The biological activity of this lot of VEGF was determined by its mitogenic activity on human umbilical vein endothelial cells (HUVEC) with an ED50 of about 1.0 ng/mL. Results may vary depending on the cell line used. VEGF will also stimulate endothelial cells from other species.
Physical form
10 µg VEGF protein was lyophilized from buffer containing 25mM HEPES and 150mM NaCl, pH 7.0. Carrier free.
10 µg of VEGF lyophilized protein from buffer containing 25mM HEPES and 150mM NaCl, pH 7.0. Carrier free.
Sterilized through a 0.2 micron membrane filter
Storage and Stability
The lyophilized powder is stable for a few weeks at room temperature, but best stored at -20°C for up to one year from date of receipt. After a quick spin, reconstitute with sterile water or 25mM HEPES and 150mM NaCl, pH 7.0. Reconstituted material can be stored at 2°-8°C for one week, but undiluted aliquots at -20°C are recommended. Avoid repeated freeze/thaw cycles.
This product is acid stable. Dilute in either 0.1% TFA or 4mM HCl, containing 25ug BSA per ug VEGF to maintain stability. Reducing reagents (such as dithiothrietol or 2-mercaptoethanol) inhibit VEGF activity.
This product is acid stable. Dilute in either 0.1% TFA or 4mM HCl, containing 25ug BSA per ug VEGF to maintain stability. Reducing reagents (such as dithiothrietol or 2-mercaptoethanol) inhibit VEGF activity.
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage Class Code
12 - Non Combustible Liquids
WGK
nwg
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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H Abedi et al.
The Journal of biological chemistry, 272(24), 15442-15451 (1997-06-13)
Vascular endothelial growth factor (VEGF) stimulated the tyrosine phosphorylation of multiple components in confluent human umbilical vein endothelial cells (HUVECs) including bands of Mr 205,000, corresponding to the VEGF receptors Flt-1 and KDR, and Mr 145,000, 120,000, 97,000, and 65,000-70,000.
Vascular endothelial cell growth factor (VEGF) produced by A-431 human epidermoid carcinoma cells and identification of VEGF membrane binding sites.
Myoken, Y, et al.
Proceedings of the National Academy of Sciences of the USA, 88, 5819-5823 (1991)
John P Morgan et al.
Nature protocols, 8(9), 1820-1836 (2013-08-31)
This protocol describes how to form a 3D cell culture with explicit, endothelialized microvessels. The approach leads to fully enclosed, perfusable vessels in a bioremodelable hydrogel (type I collagen). The protocol uses microfabrication to enable user-defined geometries of the vascular
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