Human VEGF 165 Antibody

R&D Systems | Catalog # AF-293-NA

R&D Systems
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Porcine, Avian - Chicken, Avian - Quail, Bovine, Guinea Pig, Primate - Macaca mulatta (Rhesus Macaque), Rabbit

Applications

Validated:

Immunohistochemistry, Western Blot, Neutralization, Dual RNAscope ISH-IHC Compatible, Immunocytochemistry

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Neutralization, Flow Cytometry, Immunocytochemistry, Immunoprecipitation, Affinity Assay, Bioassay, Column, ELISA Capture, ELISA Development, ELISA Development (Capture), ELISA Development (Detection), Immunoassay Development

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all VEGF Primary Antibodies »

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human VEGF165
Ala27-Arg191
Accession # NP_001165097.1

Specificity

Detects human VEGF in direct ELISAs. In direct ELISAs, approximately 100% cross-reactivity with recombinant canine VEGF,  less than 20% crossreactivity with recombinant mouse VEGF and recombinant rat VEGF, and less than 5% cross-reactivity with recombinant zebrafish VEGF is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human VEGF 165 Antibody

Detection of Human VEGF antibody by Western Blot.

Detection of Human VEGF by Western Blot.

Western blot shows lysates of human lung tissue, human kidney tissue, and human liver tissue. PVDF membrane was probed with 5-10 µg/mL of Goat Anti-Human VEGF 165Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-293-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for VEGF at approximately 27 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
VEGF antibody in HUVEC Cells by Immunocytochemistry (ICC).

VEGF in HUVEC Cells.

VEGF was detected in immersion fixed human umbilical vein endothelial cells (HUVECs) using Goat Anti-Human VEGF165 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-293-NA) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; NL001) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
VEGF antibody in Human Liver Cancer Tissue by Immunohistochemistry (IHC-P).

VEGF in Human Liver Cancer Tissue.

VEGF was detected in immersion fixed paraffin-embedded sections of human liver cancer tissue using Goat Anti-Human VEGF165 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-293-NA) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in cancer cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Cell Proliferation Induced by VEGF165and Neutralization by Human VEGF Antibody.

Cell Proliferation Induced by VEGF165and Neutralization by Human VEGF Antibody.

Recombinant Human VEGF165 (293-VE) stimulates proliferation in HUVEC human umbilical vein endothelial cells in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human VEGF165 (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human VEGF165 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-293-NA). The ND50 is typically 0.02‑0.12 µg/mL.
Detection of Human VEGF by Western Blot

Detection of Human VEGF by Western Blot

VEGF expression determined by Western blot and immunoprecipitation.A. Western blot using LiCor Odyssey to simultaneously image pan-VEGF and VEGF-A165b probed western blot. Two different podocyte samples, and a primary RPE sample were run on a gel and probed with antibodies to VEGF-A165b (mouse monoclonal anti-CTRSLTRKD, and 680nm-donkey anti-mouse, top image) and pan-VEGF (rabbit polyclonal anti-VEGF, and 800nm-donkey anti-rabbit, middle image). The bottom image is the pseudocoloured combined image (600nm green, 800nm red). Note the red VEGF165, but yellow VEGF-A165b. MWM = molecular weight marker. d = dimer, m = monomer. B. Protein extracted from human cell lines (adenoma and adenocarcinoma(AC)) subjected to immunoprecipitation (IP) for VEGF-A165b and immunoblotting (IB) for total VEGF-A. A clear strong band was seen in the IP for both cell types at ∼23kDa and ∼46kDa, consistent with the IP for recombinant human VEGF-A165b. A weaker band was seen in the input protein (not subjected to IP), and a second band slightly higher in the AC. A weak band at approximately 56kDa and 28kDa was seen in all lanes subjected to IP, including the VEGF-A165a band, but not seen in the recombinant human VEGF-A165b not subjected to IP, indicating that this is cross reactivity with the IgG. This band was clearly above the VEGF-A165b bands. C. Protein extracted from human cell lines (adenoma and adenocarcinoma(AC)) subjected to immunoprecipitation (IP) for VEGF-A and immunoblotting (IB) for VEGF-A165b. A clear strong band was seen in the IP for both cell types at ∼23kDa, the same size as recombinant human VEGF-A165b. In the input a band at ∼46Da was seen predominantly, for both cell types, labelled as VEGF-A165b dimers. D. Mouse tissues probed with VEGF-A165b antibody detect mouse IgG due to the secondary antibody. Top image, western blot of mouse tissues, recombinant mouse IgG or human VEGF-A165b or VEGF-A165b probed with mouse anti-CTRSLTRKD, and 680nm-donkey anti-mouse IgG. Bottom ima
Detection of Human VEGF by Western Blot

Detection of Human VEGF by Western Blot

Inhibitor of differentiation 4 (ID4) expression in breast cancer cells leads to the activation of an angiogenic programme in macrophages. a Expression matrix representing a panel of angiogenic factors evaluated using TaqMan Low-Density Arrays (TLDA) in macrophages obtained by 1,25-dihydroxyvitamin D3 (VitD3)-mediated differentiation of HL60 cells and subsequently cultured in RPMI medium or in conditioned media (CM) from control (EV) or ID4-overexpressing (ID4) MDA-MB-468 breast cancer cells. b Western blot showing ID4-HA overexpression in MDA-MB-468 cells. c Selected genes modulated in the arrays were evaluated by RT-qPCR in macrophages obtained from VitD3-mediated differentiation of U937 cells and subsequently cultivated in RPMI medium (CTR) or in CM from control (CM si-SCR) or ID4-depleted (CM si-ID4) MDA-MB-468 cells. d Western blot analysis showing the level of ID4 protein after transfection of the indicated small interfering RNAs (siRNAs) in MDA-MB-468 cells. e–g Western blot analysis of ephrin B2 (EphB2), granulin (GRN) and hypoxia-inducible factor (HIF)-1A proteins in differentiated U937 cells cultured in CM si-SCR or CM si-ID4 from MDA-MB-468 cells. h Immunofluorescence analysis of HIF-1A protein performed in differentiated U937 cells cultured in the presence of CM si-SCR or CM si-ID4 from MDA-MB-468 cells. i Western blotting showing the efficiency of vascular endothelial growth factor A (VEGFA) depletion by siRNA transfection in MDA-MB-468 cells used to prepare CM used in experiments shown in (j). j RT-qPCR analysis of the indicated messenger RNAs in U937 macrophages cultivated in the presence of CM from control (si-SCR) or VEGFA-depleted (si-VEGFA) MDA-MB-468 cells. k RT-qPCR analysis of the indicated genes in differentiated U937 cells cultivated in RPMI medium or in CM from MDA-MB-468 cells in the presence of VEGFA blocking antibody (Ab) or a control Ab. Specifically, VEGFA blocking Ab or control Ab were incubated with CM for 30 minutes at room temperatur
Detection of Human VEGF by Western Blot

Detection of Human VEGF by Western Blot

Western blot analysis of the effects of RNA interference on VEGF expression in HTF cells. (a) Relative expression of VEGF in cells treated with shRNA-2 and shRNA-3; (b) protein expression levels in treated groups and naïve HTF controls. ∗∗∗p < 0.01. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28168047), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of VEGF in Human Liver.

Formalin-fixed paraffin-embedded tissue sections of human liver were probed for VEGF mRNA (ACD RNAScope Probe, catalog #423168, Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using goat anti-human VEGF polyclonal antibody (R&D Systems AF-293-NA) at 5ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-goat IgG VisUCyte HRP Polymer Antibody (VC004) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to hepatocytes.
Human VEGF ELISA Standard Curve

Human VEGF ELISA Standard Curve

Recombinant Human VEGF 165 (293-VE) was serially diluted and captured by Mouse Anti-Human/Primate VEGF Monoclonal Antibody (MAB293) coated on a Clear Polystyrene Microplate (DY990). Goat Anti-Human VEGF 165 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-293-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (DY998)
Human VEGF DuoSet ELISA

Human VEGF DuoSet ELISA

Recombinant Human VEGF 165 (Catalog # 293-VE) was serially diluted and captured by Mouse Anti-Human/Primate VEGF Monoclonal Antibody (MAB293) coated on a Clear Polystyrene Microplate (DY990). Goat Anti-Human VEGF 165 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-293-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (DY998)

Applications for Human VEGF 165 Antibody

Application
Recommended Usage

Dual RNAscope ISH-IHC Compatible

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human liver

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed human umbilical vein endothelial cells

Immunohistochemistry

3-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human liver cancer tissue

Western Blot

5-10 µg/mL
Sample: Human lung tissue, human kidney tissue, and human liver tissue

Neutralization

Measured by its ability to neutralize VEGF165-induced proliferation in HUVEC human umbilical vein endothelial cells. Conn, G. et al. (1990) Proc. Natl. Acad. Sci. USA 87:1323. The Neutralization Dose (ND50) is typically 0.02‑0.12 µg/mL in the presence of 10 ng/mL Recombinant Human VEGF165.

Reviewed Applications

Read 5 reviews rated 4.6 using AF-293-NA in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.<br/>*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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. It is a member of the PDGF family that is characterized by the presence of eight conserved cysteine residues and a cystine knot structure. VEGF165 appears to be the most abundant and potent isoform, followed by VEGF121 and VEGF189. Human VEGF165 is an approximately 44 kDa molecular weight homodimer sharing 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. Although VEGF affinity is highest for binding to VEGF R1, VEGF R2 appears to be the primary mediator of VEGF angiogenic activity. VEGF165 binds the Semaphorin receptor, Neuropilin-1 and promotes complex formation with VEGF R2. VEGF is required during embryogenesis and functions to regulate the proliferation, migration, and survival of endothelial cells. In adults, VEGF functions mainly in wound healing and the female reproductive cycle. Pathologically, it is involved in tumor angiogenesis and vascular leakage. Circulating VEGF levels correlate with disease activity in autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and systemic lupus erythematosus. VEGF is induced by hypoxia and cytokines such as IL-1, IL-6, IL-8, Oncostatin M (OSM) and TNF-alpha.

Long Name

Vascular Endothelial Growth Factor

Alternate Names

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

Entrez Gene IDs

7422 (Human); 22339 (Mouse); 83785 (Rat); 281572 (Bovine); 403802 (Canine); 493845 (Feline); 30682 (Zebrafish)

Gene Symbol

VEGFA

UniProt

NP_001165097.1

Additional VEGF Products

Product Documents for Human VEGF 165 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human VEGF 165 Antibody

For research use only

Citations for Human VEGF 165 Antibody

Customer Reviews for Human VEGF 165 Antibody (5)

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Customer Images

Showing  1 - 5 of 5 reviews Showing All
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  • Human VEGF165 Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: DU145 human prostate carcinoma cell line
    Species: Human
    Verified Customer | Posted 11/12/2025
    Human VEGF 165 Antibody AF-293-NA
  • Human VEGF165 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Skeletal muscle
    Species: Human
    Verified Customer | Posted 02/25/2020
    Lane 7 shows the VEGF positive control 293-VE-010.
    Human VEGF 165 Antibody AF-293-NA
  • Human VEGF165 Antibody
    Name: Zoia Levashova
    Application: Meso Scale Discovery
    Sample Tested: Vitreous humor, Aqueous humor and Retina/choroid
    Species: Porcine
    Verified Customer | Posted 06/22/2018
    After labeling with Sulfo-Tag, used as a detection reagent in MSD assay to detect ocular VEGF level in a swine model of laser-induced choroidal neovascularization. A standard curve with pig VEGF from Bio-Rad (Cat# PPP030) is shown (2-8,000 pg/ml).
    Human VEGF 165 Antibody AF-293-NA
  • Human VEGF165 Antibody
    Name: Dong Liu
    Application: Block/Neutralize
    Sample Tested: Primary pumonary arterial endothelial cell lysate
    Species: Human
    Verified Customer | Posted 11/23/2015
    Neutralization of recombinant VEGF-induced VEGFR2 phosphorylation.
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 22897854
    Species: Human
    Verified Customer | Posted 01/07/2015

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Protocols

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FAQs for Human VEGF 165 Antibody

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  • Q: Does Human VEGF165 Antibody, Catalog # AF-293-NA, recognize VEGF189?

    A: Yes, AF-293-NA detects VEGF189

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