vascular endothelial growth factor A-The lifeline of tumor-1

VEGF-A(Vascular Endothelial Growth Factor A), belongs to platelet derived growth factor family, mainly distributed in human and animal’s kidney, spleen, liver, skeleton, brain etc. VEGFA increases vascular permeability, promotes the multiplication and migration of vascular endothelial cell, decreases the tumor apoptosis, and enhances the generation of vascular support by binding with VEGFR1, VEGFR2, Nrp1 and Nrp2.

VEGF-A mRNA consists of 8 exons and 7 introns, and yield 7 isoforms and 9 isomers by alternative splicing, mainly includes VEGF-A121, VEGF-A145, VEGF-A165, VEGF-A183, VEGF-A189 and VEGF-A206. The arabic number means the number of amino acids. The schematic diagram about their respective amino acid sequences is shown in Fig 1. All of them could regulate the lifeblood, but their locations and biological activities varies in different isoforms which relate to the amino acids, and their biological activities are associated with the expression of their sulfate-proteoglycan domains.

VEGF-A165, a basic protein which locates in both of extracellular and cytoplasmic matrix, has a heparin sulfate proteoglycan domain and possesses the optimal bioavailability and biotic potential [1]. VEGF-A121 and VEGF-A145 are weak acidic and secretory proteins, they don’t have heparin sulfate proteoglycan domain and cannot bind glycosidoprotein in extracellular matrix. The alkalescence of VEGF-A183, VEGF-A189, and VEGF-A206 are stronger than VEGF-A165, they are mainly distributed in extracellular matrix and have strong affinities to heparin.

In order to Cloud-Clone Corp. developed a series of detection tools on VEGF-A, those products include recombinant proteins, antibodies and Elisa kits. The development of VEGF-A products brings researchers great access to VEGF-A research.

By analyzing the sequence of VEGF-A, we chose Pro28~Asp191 from VEGF-A and cloned this fragment into corresponding plasmid, expressed in E.coli to get recombinant VEGF-A protein(RPA143Hu01). Regarding its molecular weight and purity, we had verified by SDS-PAGE assay, please see Fig.2.

After immunizing mice with RPA143Hu01 and we purified it by affinity chromatography to get Monoclonal Antibody to VEGF-A (MAA143Hu21); so did the Polyclonal Antibody to VEGF-A (PAA143Hu01). After coupling PAA143Hu01 with biotin, we obtained Biotin-Linked Antibody to VEGF-A (PAA143Hu71), and we also can obtain FITC-Linked Antibody to VEGF-A via coupling PAA143Hu01 with FITC (PAA143Hu81). Both these two antibodies not only can recognize VEGF-A but also have fluorescence or biotin labelling, which would be more convenient for people to detect VEGF-A in vivo and in vitro. These two antibodies have been tested with Western Blot, IHC and so on. From theWestern Blotting result in Fig.3, PAA143Hu01 can specifically recognize the VEGF-A in the biological samples. Furthermore, the IHC result shows that the VEGF-A mainly distributed in the cytoplasm of hepatocytes (Fig.4).

Based on the recombinant protein and antibodies, Cloud-Clone Corp. developed VEGFA kits, including Elisa kit (SEA143Hu) and CLIA kit (SCA143Hu).

Further, Cloud-Clone Corp. developed over 40 VEGFA-related products against VEGFA that originated from rat, mouse, rabbit, and so on, Those products include protein, antibody, biotin conjugated antibody, and FITC conjugated antibody, Elisa kit and CLIA kit. These products are great access to investigate the function of VEGF-A for researchers.

Last but not the least, in order to investigate specific VEGF-A isoforms, Cloud-Clone Corp. also developed products against other VEGF-A isoforms, including VEGF-A121, VEGF-A145, VEGF-A165, VEGF-A183, VEGF-A189 and VEGF-A206 . Those products relates to protein, antibody, biotin conjugated antibody, FITC conjugated antibody, Elisa kit, CLIA kit, etc. During the process of development, Cloud-Clone Corp. screened a large number of antibodies to avoid the cross-reactivities among different isoforms. So the kits can detect target protein specificially, while other isoforms won’t interfere the detection.

For more information, please visit http://www.cloud-clone.us/.

[1]Ferrara Napoleone,Gerber Hans Peter. Vascular endothelial growth factor molecular and biological aspects [J]. Advances in Organ Biology, 1999, 7(25-57).