Overview

  • Product nameAnti-VEGFA antibody
    See all VEGFA primary antibodies
  • Description
    Rabbit polyclonal to VEGFA
  • SpecificityThis antibody is detecting different isoforms of VEGF between 15-40 kDa. For Isoforms see http://www.uniprot.org/uniprot/P15692#sequences
  • Tested applicationsSuitable for: ICC/IF, WB, IHC-Fr, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human VEGFA aa 50-150 conjugated to Keyhole Limpet Haemocyanin (KLH).
    (Peptide available as ab46161)

  • Positive control
    • Purchase matching WB positive control:Recombinant Human VEGFA protein
    • ab46154 gave a positive result in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) A431 (Human epithelial carcinoma cell line) HEK 293 (Human embryonic kidney cell line) HepG2 (Human hepatocellular liver carcinoma cell line) MCF7 (Human breast adenocarcinoma cell line) SHSY-5Y (Human neuroblastoma cell line) IHC: Human Cerebellum (FFPE sections) ICC/IF: SV40LT-SMC cells

Properties

Applications

Our Abpromise guarantee covers the use of ab46154 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 43 kDa (predicted molecular weight: 24, 45 kDa).
IHC-Fr 1/200.
IHC-P Use a concentration of 5 - 10 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • FunctionGrowth factor active in angiogenesis, vasculogenesis and endothelial cell growth. Induces endothelial cell proliferation, promotes cell migration, inhibits apoptosis and induces permeabilization of blood vessels. Binds to the FLT1/VEGFR1 and KDR/VEGFR2 receptors, heparan sulfate and heparin. NRP1/Neuropilin-1 binds isoforms VEGF-165 and VEGF-145. Isoform VEGF165B binds to KDR but does not activate downstream signaling pathways, does not activate angiogenesis and inhibits tumor growth.
  • Tissue specificityIsoform VEGF189, isoform VEGF165 and isoform VEGF121 are widely expressed. Isoform VEGF206 and isoform VEGF145 are not widely expressed.
  • Involvement in diseaseDefects in VEGFA are a cause of susceptibility to microvascular complications of diabetes type 1 (MVCD1) [MIM:603933]. These are pathological conditions that develop in numerous tissues and organs as a consequence of diabetes mellitus. They include diabetic retinopathy, diabetic nephropathy leading to end-stage renal disease, and diabetic neuropathy. Diabetic retinopathy remains the major cause of new-onset blindness among diabetic adults. It is characterized by vascular permeability and increased tissue ischemia and angiogenesis.
  • Sequence similaritiesBelongs to the PDGF/VEGF growth factor family.
  • Cellular localizationSecreted. VEGF121 is acidic and freely secreted. VEGF165 is more basic, has heparin-binding properties and, although a signicant proportion remains cell-associated, most is freely secreted. VEGF189 is very basic, it is cell-associated after secretion and is bound avidly by heparin and the extracellular matrix, although it may be released as a soluble form by heparin, heparinase or plasmin.
  • Information by UniProt
  • Database links
  • Alternative names
    • Folliculostellate cell-derived growth factor antibody
    • Glioma-derived endothelial cell mitogen antibody
    • MGC70609 antibody
    • MVCD1 antibody
    • Vascular endothelial growth factor A antibody
    • vascular endothelial growth factor A121 antibody
    • vascular endothelial growth factor A165 antibody
    • vascular endothelial growth factor antibody
    • Vascular permeability factor antibody
    • VEGF A antibody
    • Vegf antibody
    • VEGF-A antibody
    • VEGF120 antibody
    • Vegfa antibody
    • VEGFA_HUMAN antibody
    • VPF antibody
    see all

Anti-VEGFA antibody images

  • IHC image of ab46154 staining VEGF in human cerebellum formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab46154, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

     

  • ab46154 staining VEGF in SV40LT-SMC cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab46154 at a working concentration of 5μg/ml and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-rabbit AlexaFluor® 488 (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • All lanes : Anti-VEGFA antibody (ab46154) at 1 µg

    Lane 1 : MEF whole cell lysate
    Lane 2 : HeLa whole cell lysate
    Lane 3 : MCF7 whole cell lysate
    Lane 4 : NIH 3T3 whole cell lysate
    Lane 5 : RCC4 whole cell lysate
    Lane 6 : PC12 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat anti-rabbit IgG (IRDye 800) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 24, 45 kDa
  • All lanes : Anti-VEGFA antibody (ab46154) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 24, 45 kDa
    Observed band size : 42 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab46154 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

  • ab46154 staining VEGF in HeLa cells treated with Y-27632 dihydrochloride (ab120129), by ICC/IF. Decrease in VEGF expression correlates with increased concentration of Y-27632 dihydrochloride, as described in literature.
    The cells were incubated at 37°C for 1h in media containing different concentrations of ab120129 (Y-27632 dihydrochloride) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab46154 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • All lanes : Anti-VEGFA antibody (ab46154) at 0.5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : A431 whole cell lysate (ab7909)
    Lane 4 : HEK293 whole cell lysate (ab7902)
    Lane 5 : HepG2 whole cell lysate (ab7900)
    Lane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
    Lane 8 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with VEGFA peptide (ab46160) at 1 µg/ml
    Lane 9 : Jurkat whole cell lysate (ab7899) with VEGFA peptide (ab46160) at 1 µg/ml
    Lane 10 : A431 whole cell lysate (ab7909) with VEGFA peptide (ab46160) at 1 µg/ml
    Lane 11 : HEK293 whole cell lysate (ab7902) with VEGFA peptide (ab46160) at 1 µg/ml
    Lane 12 : HepG2 whole cell lysate (ab7900) with VEGFA peptide (ab46160) at 1 µg/ml
    Lane 13 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate with VEGFA peptide (ab46160) at 1 µg/ml
    Lane 14 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate with VEGFA peptide (ab46160) at 1 µg/ml

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 24, 45 kDa
  • ICC/IF image of this anti-VEGF antibody (ab46154) stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab46154, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • ab46154 staining kidney tissue sections (human, mouse and rat) by IHC-P.  Sections were formaldehyde fixed and subjected to heat mediated antigen retrieval prior to blocking with sequential peroxidase and protein block (prediluted) for 20 minutes at 20°C.  The anti-VEGF antibody was diluted 1/100 and incubated with the samples for 45 minutes at 20°C.  A HRP-conjugated goat anti-rabbit antibody was used as the secondary.

    See Abreview

  • ab46154 staining VEGF in MCF7 cells treated with HA 1077 (ab120306), by ICC/IF. Decrease in VEGF expression correlates with increased concentration of HA 1077 as described in literature.
    The cells were incubated at 37°C for 1h in media containing different concentrations of ab120306 (HA 1077) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab46154 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody.
  • IHC image of VEGF staining in human heart formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab46154, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • ICC/IF image of ab46154 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab46154, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-VEGFA antibody (ab46154)

This product has been referenced in:
  • Chen X  et al. Suppression of HSP27 increases the anti-tumor effects of quercetin in human leukemia U937 cells. Mol Med Rep 13:689-96 (2016). WB ; Human . Read more (PubMed: 26648539) »
  • Soriano A  et al. MicroRNA-497 impairs the growth of chemoresistant neuroblastoma cells by targeting cell cycle, survival and vascular permeability genes. Oncotarget 7:9271-87 (2016). WB . Read more (PubMed: 26824183) »

See all 130 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (MCF-7)
Gel Running Conditions Reduced Denaturing
Loading amount 20 µg
Specification MCF-7
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 29 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Prostate)
Antigen retrieval step None
Permeabilization No
Specification Prostate
Blocking step Serum Free Protein Block (Dako) as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 24°C
Fixative 10% NBF
Username

Ms. Premier Laboratory

Verified customer

Submitted May 18 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Human Synoviocyte fibroblast-like cells)
Gel Running Conditions Reduced Denaturing (8% SDS-PAGE)
Loading amount 35 µg
Treatment High glucose and Palmitic acid for 24hrs
Specification Human Synoviocyte fibroblast-like cells
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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Submitted Apr 19 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (GONAD)
Antigen retrieval step Enzymatic
Permeabilization No
Specification GONAD
Blocking step Serum as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Paraformaldehyde
Username

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Verified customer

Submitted Jun 22 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (SKOV3 ovarian carcinoma cells)
Permeabilization Yes - 0.25% Triton X-100 in PBS
Specification SKOV3 ovarian carcinoma cells
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Paraformaldehyde
Username

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Verified customer

Submitted May 06 2015

Application Western blot
Sample Sheep Tissue lysate - whole (lung)
Gel Running Conditions Reduced Denaturing (12%)
Loading amount 15 µg
Specification lung
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Miss. Maryline Gomes

Verified customer

Submitted Feb 13 2015

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (Hela, Hep G2)
Specification Hela, Hep G2
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Jan 02 2015

Thank you for your enquiry.
There are many VEGFA isoforms
http://www.uniprot.org/uniprot/P15692#P15692
The immunogen for ab46154 VEGF antibody has 100% alignment to all the VEGFA isoforms described on this protein database page and so would...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Sample Mouse Tissue sections (Heart 3 valve area)
Specification Heart 3 valve area
Permeabilization No
Fixative Formaldehyde
Username

Ms. B Ren

Verified customer

Submitted Aug 30 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Breast cacner)
Loading amount 20 µg
Specification Breast cacner
Gel Running Conditions Reduced Denaturing (12% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted May 10 2013

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