概述

  • 产品名称
  • 描述
    兔多克隆抗体to VEGFA
  • 特异性
    Recent batches of this product have not passed our stringent batch testing criteria in ICC/IF and therefore ab46154 is no longer guaranteed in this application. Please contact customer support with any batch specific queries. This antibody is detecting different isoforms of VEGF between 15-40 kDa. For Isoforms see http://www.uniprot.org/uniprot/P15692#sequences
  • 经测试应用
    适用于: WB, IHC-Fr, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

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

  • 阳性对照
    • 购买相配的WB阳性对照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)

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能
    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.
  • 组织特异性
    Isoform VEGF189, isoform VEGF165 and isoform VEGF121 are widely expressed. Isoform VEGF206 and isoform VEGF145 are not widely expressed.
  • 疾病相关
    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.
  • 序列相似性
    Belongs to the PDGF/VEGF growth factor family.
  • 细胞定位
    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
  • 数据库链接
  • 别名
    • 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 图像

  • 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.

     

  • 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

  • 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
  • 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

  • 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.

Anti-VEGFA antibody (ab46154)参考文献

This product has been referenced in:
  • Lee A  et al. Hair growth promoting effect of dermal papilla like tissues from canine adipose-derived mesenchymal stem cells through vascular endothelial growth factor. J Vet Med Sci 78:1811-1818 (2017). IHC, WB, IF ; Dog . Read more (PubMed: 27647656) »
  • Solà-Adell C  et al. Calcium Dobesilate Prevents Neurodegeneration and Vascular Leakage in Experimental Diabetes. Curr Eye Res N/A:1-14 (2017). Read more (PubMed: 28574750) »

See all 138 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

提交于 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
Username

Miss. Seulgee Lee

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 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

Premier Laboratory

Verified customer

提交于 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

Abcam user community

Verified customer

提交于 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

Abcam user community

Verified customer

提交于 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

Abcam user community

Verified customer

提交于 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

提交于 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
Username

Abcam user community

Verified customer

提交于 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...

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