Overview

  • Product name
  • Description
    Rabbit polyclonal to VEGFA
  • Host species
    Rabbit
  • Specificity
    New batches of this antibody are no longer guaranteed in ICC/IF, IHC-P or IHC-Fr as they have not passed our stringent batch testing criteria. Please contact customer support for any specific queries. We recommend ab52917 as an alternative for ICC/IF and IHC-P.
  • Tested applications
    Suitable for: WB, ELISAmore 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

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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 27 kDa).

We recommend Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody.

ELISA Use at an assay dependent concentration.

Target

  • Function
    Growth 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 specificity
    Isoform VEGF189, isoform VEGF165 and isoform VEGF121 are widely expressed. Isoform VEGF206 and isoform VEGF145 are not widely expressed.
  • Involvement in disease
    Defects 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 similarities
    Belongs to the PDGF/VEGF growth factor family.
  • Cellular localization
    Secreted. 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

Images

  • Anti-VEGFA antibody (ab46154) at 1 µg/ml + HCT116 whole cell lysate at 20 µg

    Secondary
    Rabbit IgG secondary antibody at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 27 kDa
    Observed band size: 23 kDa
    why is the actual band size different from the predicted?


    Exposure time: 2 minutes


    This blot was produced using a 10% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin 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.

  • Ab46154 was tested using indirect ELISA. The wells were coated with peptide (1 µg x mL-1 at 100 µL per well) overnight at 4°C, followed by a 1% fat-free milk blocking step for 1 hour at room temperature. The primary antibody (ab46154) was added at a range of dilutions (50 µL per well) for 1 hour at room temperature. Ab97080 (Goat Anti-Rabbit IgG H&L-HRP) was used as a secondary antibody at 1:50.000 dilution for 1 hour at room temperature and signal was developed with TMB substrate.

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

    Lane 1 : Recombinant Human VEGFA protein (ab204773)
    Lane 2 : Recombinant mouse VEGFA protein (Active) (ab185265)

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    All lanes : Rabbit IgG secondary antibody at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 27 kDa
    Observed band size: 23 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    This blot was produced using a 10% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 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.

References

This product has been referenced in:
  • Zou X  et al. Endogenous hormone 2-methoxyestradiol suppresses venous hypertension-induced angiogenesis through up- and down-regulating p53 and id-1. J Cell Mol Med 22:957-967 (2018). Read more (PubMed: 29193609) »
  • Nürnberg C  et al. Dataset on the activation of Müller cells through macrophages upon hypoxia in the retina. Data Brief 16:489-500 (2018). WB ; Dog . Read more (PubMed: 29255783) »
See all 228 Publications for this product

Customer reviews and Q&As

1-10 of 44 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (footpad)
Permeabilization
Yes - triton
Specification
footpad
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Dr. Tina Yuan

Verified customer

Submitted Sep 10 2018

Abreviews
Application
Western blot
Sample
Mouse Cell lysate - whole cell (EMT6 cell line)
Gel Running Conditions
Reduced Denaturing (4-20% Tris Glycin gel, semi dry transfer)
Loading amount
50 µg
Specification
EMT6 cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jul 17 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (HCT 116 cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Specification
HCT 116 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jul 17 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Vascular smooth muscle cell)
Permeabilization
Yes - NP40
Specification
Vascular smooth muscle cell
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jul 27 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (vessel)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
No
Specification
vessel
Blocking step
dako antibody diluent with background reducing components as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1µg/mL · Temperature: 25°C
Fixative
Formaldehyde

Miss. Seulgee Lee

Verified customer

Submitted Feb 21 2017

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

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

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

Abcam user community

Verified customer

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

Abcam user community

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

Abcam user community

Verified customer

Submitted May 06 2015

1-10 of 44 Abreviews or Q&A

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