CyTOF-ready, ELISA, Flow Cytometry, Immunocytochemistry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG₁ kappa Clone # VG1

Format

Azide and BSA Free

Concentration

1 mg/ml

View all VEGF Antibodies »

Product Specifications

Immunogen

Recombinant VEGF 189 protein.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁ kappa

Scientific Data Images for VEGF Antibody (VG1) - Azide and BSA Free

Western Blot Analysis of VEGF in Transfected Cell Lines

HUVEC and 293T cells transfected with a plasmid expressing human VEGF165 at 1:1000. Image provided by verified customer review. Image from the standard format of this antibody.

Immunohistochemical Analysis of VEGF in Paraffin Embedded Human Angiosarcoma Tissue

FFPE human angiosarcoma tissue section using VEGF antibody (clone VG1). The endothelial cells of the blood vessels and most of the cancer cells showed strong positivity for VEGF protein. Image from the standard format of this antibody.

Flow Cytometry of HUVEC Cells Stained with PerCP Conjugated VEGF Antibody

Analysis of PerCP conjugate of NB100-664. An intracellular stain was performed on HUVEC cells with VEGF (VG1) antibody NB100-664PCP (blue) and a matched isotype control NBP2-27287PCP (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were directly conjugated to PerCP.

Western Blot Analysis of VEGF in Human Kidney

Analysis of VEGF in human kidney protein using NB100-664. Image from the standard format of this antibody.

Immunocytochemistry/Immunofluorescence Staining of VEGF in HeLa Cells

HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-VEGF (VG1) [NB100-664] at 1:200 overnight at 4C and detected with an anti-mouse DyLight 488 (Green) at 1:500. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective. Image from the standard format of this antibody.

Immunocytochemistry/Immunofluorescence Staining of VEGF in MCF7 Cells

MCF7 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with VEGF (VG1) NB100-664 at 1:200 overnight at 4C and detected with DyLight 488 (Green). Actin was detected with Phalloidin 568 (Red) at 1:200. Nuclei were detected with DAPI (Blue). Cells were imaged using a 40X objective. Image from the standard format of this antibody.

Flow Cytometry of U-87 Cells Stained with Alexa Fluor 488 Conjugated VEGF Antibody

An intracellular stain was performed on U-87 cells with VEGF [VG1] Antibody NB100-664AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were directly conjugated to Alexa Fluor 488.

Flow Cytometry of U-87 Cells Stained with Allophycocyanin Conjugated VEGF Antibody

An intracellular stain was performed on U-87 cells with VEGF [VG1] Antibody NB100-664APC (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were directly conjugated to Allophycocyanin.

Flow Cytometry of U-937 Cells Stained with FITC Conjugated VEGF Antibody

An intracellular stain was performed on U-937 cells with NB100-664F (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were directly conjugated to FITC.

Flow Cytometry of U937 Cells Stained with Allophycocyanin Conjugated VEGF Antibody

An intracellular stain was performed on U937 cells with VEGF [VG1] Antibody NB100-664APC (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were directly conjugated to Allophycocyanin.

Flow Cytometry of U-937 Cells Stained with VEGF Antibody

An intracellular stain was performed on U-937 cells with VEGF Antibody (VG1) NB100-664 and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody (R&D Systems, F0102B). Image from the standard format of this antibody.

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Applications for VEGF Antibody (VG1) - Azide and BSA Free

Application

Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:10 - 1:500

Immunohistochemistry

1:20 - 1:100

Immunohistochemistry-Frozen

1:20 - 1:100

Immunohistochemistry-Paraffin

1:20 - 1:100

Western Blot

1 - 2 ug/ml

Application Notes

In IHC a dilution of 1:20-1:50 was used in an ABC method. However, depending on the staining conditions employed, we suggest that the final dilution should be determined by the user. We suggest an incubation period of 30-60 minutes at room temperature. High temperature treatment of formalin-fixed tissue sections using 1mM EDTA, pH 8.0 must be performed prior to the immunostaining. This antibody is CyTOF ready.

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: VEGF

Vascular endothelial growth factor (VEGF), also called VEGF-A and vascular permeability factor (VPF), is a secreted homodimeric glycoprotein belonging to the VEGF family with a role in stimulating angiogenesis and vasculogenesis (1,2). More specifically, VEGF-A secretion from most cell types contributes to promoting endothelial cell proliferation and migration, inhibiting apoptosis, increasing vascular permeability, and wound healing (1). The VEGF family consists of several members including VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, VEGF-F, and placenta growth factor (PLGF) (1-4). As a result of alternative splicing of the eight exon VEGFA gene, there are several VEGF-A protein isoforms of 121, 145, 165, 183, 189, and 206 amino acids (aa) in length, with VEGF121 and VEGF165 being the two most expressed isoforms (1,5). Full length VEGF-A monomer has a 26 aa signal sequence plus a 206 aa (VEGF206) sequence, with a theoretic molecular weight (MW) of 27 kDa, containing VEGF receptor 1 (VEGFR1) and VEGR2 binding sites and heparin-binding domains (1-3,5,6). VEGF121 lacks heparin affinity and binds the receptor tyrosine kinases (RTKs) VEGFR1 and VEGFR2, whereas VEGF165 has moderate affinity for heparin and, in addition to being a ligand for VEGFR1 and VEGFR2, can also bind the co-receptors neuropilin 1 (NRP1) and NRP2 (1,5). Hypoxia and hypoxia-related genes such as HIF-1, EGF, and PDGF are major regulators angiogenesis and VEGF expression (1,3). VEGF signaling initiated by ligand binding to its receptors results in activation of different pathways including PI3K and MAPK and ultimately guides endothelial cell proliferation, migration, and survival (1,3). While VEGF plays an important role in promoting normal angiogenesis and blood vessel formation, its expression is often upregulated in tumors and other angiogenesis-related pathologies like osteroarthritis (OA) (1-5,7). Given its function, VEGF and its receptors have become a therapeutic target for treating cancer and blocking angiogenesis (4,5,7). A recombinant humanized monoclonal anti-VEGFA antibody called bevacizumab (Avastin) was first approved by the FDA in 2004 for the treatment of a number of cancers (1-3,5). Cancer patients may experience resistance to anti-VEGF antibodies and, as such, clinical studies are exploring combination treatment options with chemotherapies and immune-checkpoint inhibitors (3,5).

References

1. Melincovici CS, Bosca AB, susman S, et al. Vascular endothelial growth factor (VEGF) - key factor in normal and pathological angiogenesis. Rom J Morphol Embryol. 2018;59(2):455-467.

2. Shaik F, Cuthbert GA, Homer-Vanniasinkam S, Muench SP, Ponnambalam S, Harrison MA. Structural Basis for Vascular Endothelial Growth Factor Receptor Activation and Implications for Disease Therapy. Biomolecules. 2020;10(12):1673. https://doi.org/10.3390/biom10121673

3. Apte RS, Chen DS, Ferrara N. VEGF in Signaling and Disease: Beyond Discovery and Development. Cell. 2019;176(6):1248-1264. https://doi.org/10.1016/j.cell.2019.01.021

4. Matsumoto K, Ema M. Roles of VEGF-A signalling in development, regeneration, and tumours. J Biochem. 2014;156(1):1-10. https://doi.org/10.1093/jb/mvu031

5. Itatani Y, Kawada K, Yamamoto T, Sakai Y. Resistance to Anti-Angiogenic Therapy in Cancer-Alterations to Anti-VEGF Pathway. Int J Mol Sci. 2018;19(4):1232. Published 2018 Apr 18. doi:10.3390/ijms19041232

6. Uniprot (P15692)

7. Hamilton JL, Nagao M, Levine BR, Chen D, Olsen BR, Im HJ. Targeting VEGF and Its Receptors for the Treatment of Osteoarthritis and Associated Pain. J Bone Miner Res. 2016;31(5):911-924. https://doi.org/10.1002/jbmr.2828

Long Name

Vascular Endothelial Growth Factor

Alternate Names

MVCD1, VAS, Vasculotropin, VEGF-A, VEGFA, VPF

Gene Symbol

VEGFA

Additional VEGF Products

Product Documents for VEGF Antibody (VG1) - Azide and BSA Free

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for VEGF Antibody (VG1) - Azide and BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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VEGF Antibody (VG1) - Azide and BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-81007

Key Product Details

Species Reactivity

Applications

Label

Antibody Source

Format

Concentration

View all VEGF Antibodies »

Product Specifications

Immunogen

Clonality

Host

Isotype

Scientific Data Images for VEGF Antibody (VG1) - Azide and BSA Free

Western Blot Analysis of VEGF in Transfected Cell Lines

Immunohistochemical Analysis of VEGF in Paraffin Embedded Human Angiosarcoma Tissue

Flow Cytometry of HUVEC Cells Stained with PerCP Conjugated VEGF Antibody

Western Blot Analysis of VEGF in Human Kidney

Immunocytochemistry/Immunofluorescence Staining of VEGF in HeLa Cells

Immunocytochemistry/Immunofluorescence Staining of VEGF in MCF7 Cells

Flow Cytometry of U-87 Cells Stained with Alexa Fluor 488 Conjugated VEGF Antibody

Flow Cytometry of U-87 Cells Stained with Allophycocyanin Conjugated VEGF Antibody

Flow Cytometry of U-937 Cells Stained with FITC Conjugated VEGF Antibody

Flow Cytometry of U937 Cells Stained with Allophycocyanin Conjugated VEGF Antibody

Flow Cytometry of U-937 Cells Stained with VEGF Antibody

Applications for VEGF Antibody (VG1) - Azide and BSA Free

Immunocytochemistry/ Immunofluorescence

Immunohistochemistry

Immunohistochemistry-Frozen

Immunohistochemistry-Paraffin

Western Blot

Formulation, Preparation, and Storage

Purification

Formulation

Format

Preservative

Concentration

Shipping

Stability & Storage

Background: VEGF

Long Name

Alternate Names

Gene Symbol

Additional VEGF Products

Product Documents for VEGF Antibody (VG1) - Azide and BSA Free

Product Datasheets

COA

Certificate of Analysis

SDS

Product Specific Notices for VEGF Antibody (VG1) - Azide and BSA Free