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Sigma-Aldrich

VEGF Protein, Human recombinant

The human recombinant VEGF Protein is available in a 10 µg format.

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UNSPSC Code:
12352202
eCl@ss:
32160405
NACRES:
NA.75

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)

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 37C 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.

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.

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.

wgk_germany

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.
Vishnu Suresh Babu et al.
Cells, 11(20) (2022-10-28)
Mutations in the RB1 locus leading to a loss of functional Rb protein cause intraocular tumors, which uniquely affect children worldwide. These tumors demonstrate rapid proliferation, which has recently been shown to be associated with an altered metabolic signature. We
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
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)
Elisa Pardella et al.
Cells, 11(24) (2022-12-24)
Cancer progression is supported by the cross-talk between tumor cells and the surrounding stroma. In this context, senescent cells in the tumor microenvironment contribute to the development of a pro-inflammatory milieu and the acquisition of aggressive traits by cancer cells.

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