製品の概要

  • 製品名
    Anti-VEGFA antibody [VG-1]
    VEGFA 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [VG-1] to VEGFA
  • 特異性
    Detects the 121, 165 and 189 VEGF isoforms in routinely fixed specimens.
  • アプリケーション
    適用あり: ICC/IF, IP, WB, ELISA, ICC, IHC-P, IHC-Frmore details
  • 種交差性
    交差種: Mouse, Rat, Rabbit, Dog, Human
    非交差種: Cow
  • 免疫原

    BALB/C mice were injected with a recombinant VEGF protein.

  • ポジティブ・コントロール
  • 特記事項

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab1316 in the following tested applications.

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

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 5 µg/ml.
IP 1/1000.

See Abreview.

WB Use a concentration of 5 - 10 µg/ml.

Please note in our hands this antibody is sensitive to blocking conditions. If you are seeing no bands on your western blot, we suggest 3% milk blocking with 1 hr primary incubation step at RT rather than overnight at 4°C. We would also recommend incubating primary and secondary in TBST only for this antibody.

ELISA Use at an assay dependent concentration.
ICC Use a concentration of 10 µg/ml. See Abreview.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.

ターゲット情報

  • 機能
    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.
  • 組織特異性
    Isoform VEGF189, isoform VEGF165 and isoform VEGF121 are widely expressed. Isoform VEGF206 and isoform VEGF145 are not widely expressed.
  • 関連疾患
    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.
  • 配列類似性
    Belongs to the PDGF/VEGF growth factor family.
  • 細胞内局在
    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
  • 参照データベース
  • 別名
    • 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

画像

  • All lanes : Anti-VEGFA antibody [VG-1] (ab1316)

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

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    Mouse IgG secondary antibody at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 24, 45 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 ab1316 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.

  • IHC image of ab1316 staining VEGF in rat 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 ab1316, 5μg/ml working concentration, 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.

  • ab1316 at 10µg/ml staining human retinal pigment epithelial cells. The antibody was incubated with the cells for 6 hours and then detected with Alexa-Fluor ® 568 goat anti-mouse (IgG) antibody.

    This image is courtesy of an Abreview submitted on 16 November 2005.

    See Abreview

  • IHC image of ab1316 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 ab1316, 5μg/ml working concentration, 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.

  • ICC/IF image of ab1316 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab1316, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 100% methanol fixed (5 min) Hek293, HepG2 and MCF7 cells at 10µg/ml, and in 4% PFA fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 10µg/ml.
  • ab1316 staining VEGF in Human MDA-MD-231 cells injected into the mouse mammary fat pad by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized with Tween in PBS and blocked with 1.5% serum for 1 hour at 25°C; antigen retrieval was by heat mediation in citrate buffer. Tissue samples were incubated with primary antibody (1/200 in PBST +1% BSA) for 16 hours at 4°C. A biotin-conjugated Goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody. Tissue was counterstained with Hematoxylin (1/10) for 30 seconds at room temperature and rinsed with water.

    See Abreview

  • IHC image of ab1316 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 ab1316, 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.

    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.

参考文献

This product has been referenced in:
  • Luo M  et al. Construction and expression of a lentivirus expression vector carrying the VEGF165-EGFP fusion gene in breast cancer MCF-7 cells. Oncol Lett 13:1745-1752 (2017). WB . Read more (PubMed: 28454319) »
  • Zhang HT  et al. Early VEGF inhibition attenuates blood-brain barrier disruption in ischemic rat brains by regulating the expression of MMPs. Mol Med Rep 15:57-64 (2017). IHC ; Rat . Read more (PubMed: 27909732) »

See all 71 Publications for this product

レビューと Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (293T, HepG2, MEF)
Loading amount
20 µg
Specification
293T, HepG2, MEF
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Username

Mr. Dongil Kim

Verified customer

投稿 Feb 01 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tonsil)
Specification
Tonsil
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrat buffer pH 6
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 Aug 03 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Purified protein (culture medium from astrocytes)
Loading amount
10 µg
Specification
culture medium from astrocytes
Gel Running Conditions
Non-reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Jun 23 2010

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Cat Cell lysate - whole cell (Skin Fibroblasts and Vaccination Associated Sarcom)
Gel Running Conditions
Reduced Denaturing (12% Gel)
Loading amount
10 µg
Specification
Skin Fibroblasts and Vaccination Associated Sarcom
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Username

Abcam user community

Verified customer

投稿 Nov 06 2017

いつも大変お世話になっております。アブカム株式会社の井口でございます。


この度はお問合せいただきまして誠にありがとうございます。




こちらの製品をご紹介申し上げます。


Anti-VEGFA antibody [VG-1] (ab1316)


Mouse monoclonal


Application: ELISA, ICC, ICC/IF, IHC...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (brain)
Permeabilization
Yes - Triton X-100
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Miss. Tamara Martínez Valverde

Verified customer

投稿 Dec 07 2015

Application
Other
Sample
Human Purified protein (HEK293)
Username

Abcam user community

Verified customer

投稿 Aug 19 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Antigen retrieval step
None
Sample
Rabbit Tissue sections (skin)
Specification
skin
Permeabilization
No
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

投稿 Jun 02 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric
Sample
Rabbit Tissue sections (Periosteum of rabbit)
Specification
Periosteum of rabbit
Permeabilization
No
Fixative
Paraformaldehyde
Username

Dr. 金录 郭

Verified customer

投稿 Jan 07 2014

The Active human VEGF full length protein (ab9571) has been tested on HUVECs and is known to stimulate proliferation of these cells, which shows that the GAG sites are present and active.
The recombinant human VEGF165 is a 38.2 kDa disulfide-linke...

Read More

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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