Overview

  • Product name
    Anti-VEGFA antibody [VG-1]
    See all VEGFA primary antibodies
  • Description
    Mouse monoclonal [VG-1] to VEGFA
  • Host species
    Mouse
  • Specificity
    Detects the 121, 165 and 189 VEGF isoforms in routinely fixed specimens.
  • Tested applications
    Suitable for: ICC/IF, IP, WB, ELISA, ICC, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Dog, Human
    Does not react with: Cow
  • Immunogen

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

  • Positive control
  • General notes

    Western blot protocol advice: 
    Please note that expression of VEGFA is low in most samples without treatment (e.g. hypoxia / DFX / CoCl2). Cell treatment with a Golgi inhibitor (e.g. brefeldin A) may enhance detection. Loading a high amount of sample (>50 µg) and addition of protease inhibitors (e.g. ab65621) may also enhance detection. 

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    Abcam recommended secondaries - Goat Anti-Mouse HRP (ab205719) and Goat Anti-Mouse Alexa Fluor® 488 (ab150113).

Properties

Applications

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.

Application Abreviews Notes
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 using 3% milk for 1 hr at RT for blocking. For the primary antibody incubation step, we suggest 1 hr at RT rather than overnight at 4°C. We would also recommend incubating primary and secondary antibodies in TBST only.

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.

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

  • 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.

  • 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 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.
  • 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

  • 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.
  • 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
    All lanes : 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.

References

This product has been referenced in:
  • Ma J  et al. Effect of 410 nm photodynamic therapy with hemoporfin on the expression of vascular endothelial growth factor (VEGF) in cultured human vascular endothelial cells. Lasers Med Sci 34:149-155 (2019). Read more (PubMed: 30350123) »
  • Fu YC & Xin ZM Inhibited corneal neovascularization in rabbits following corneal alkali burn by double-target interference for VEGF and HIF-1a. Biosci Rep 39:N/A (2019). Read more (PubMed: 30355648) »
See all 110 Publications for this product

Customer reviews and Q&As

1-10 of 24 Q&A

Answer

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-Fr, IHC-P, IP, WB


Reactivity: Mouse, Rat, Rabbit, Dog, Human


100 µg¥73,000 国内在庫ございます


https://www.abcam.co.jp/vegfa-antibody-vg-1-ab1316.html 




追加のご質問等ございましたら、お知らせください。

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Answer

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-linked homodimeric protein consisting of two 165 amino acid polypeptide chains. Unfortunately, we cannot be certain on where exactly the GAG sites are present. However, because ab9571 has been tested on HUVECs and does stimulate proliferation we would think that the GAG sites are present and active. Please be aware that anyhow we cannot guarantee it.
The Anti-VEGF antibody [VG-1] (ab1316) is able to detect Active human VEGF full length protein (ab9571) because ab9571 is VEGF 165 and ab1316 is able to detect VEGF 165 (please refer to the antibody datasheet for more information).

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Answer

Thank you for contacting us.


I will gladly help you to find a suitable antibody for your experiment. However, it seems that both targets are likely to produce an abundance of side bands in WB as both occur in many isoforms. In addition, even when we may have a lot of antibodies for the targets, it looks like HepG2 cells have not been used in many cases (especially with the BRCA1 target). In order to distinguish the signal from the side bands, I would like to suggest to tag the protein or to run a positive control in form of the recombinant proteins in order to check how well the antibody is picking up the signal.

The two antibodies I have picked are the following

Click here (or use the following: https://www.abcam.com/index.html?datasheet=1316). ab1316 Anti-VEGF antibody [VG-1] has been used with HepG2 cells, and has received good customer rating.

https://www.abcam.com/index.html?datasheet=79401 (or use the following: https://www.abcam.com/index.html?datasheet=79401).

ab79401 Anti-BRCA1 antibodyseems to produce a nice clear signal.



I hope this information is helpful to you. Please let me know if these two antibodies are suitable for you.
Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for contacting us. To our knowledge, the epitope for the VG-1 clone has not been mapped. While we cannot say for certain which isoforms will be recognized in rat, bands are observed at ˜27 and 31 kDa in rat dermal epithelium samples. I hope this helps, please let me know if you need any additional information or assistance.

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Answer

Thank you for your answer.

I have made now an alignment of the sequence you provided,with the three sequences we know the ab1316 does react with. Indeed the sequences are in identical in the the main parts. Please see here by yourself:
hhttp://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20120410-153045-0967-732581-pg

I can therefore confirm that the epitope is contained in your recombinantly expressed protein.
The antibody ab1316 will therefore most likely recognise your recombinantly expressed protein, if the expression is correct.

I hope this information is helpful. Please do not hesitate to contact us again should you have any concern or question.

Read More

Answer

Thank you for your inquiry.

We have not determined the exact epitope of the antibody ab1316. I can however confirm that this antibody does recognize the human121, 165 and 189 VEGF isoforms (SwissProt: P15692) and I would expect therefore that it would recognize also the recombinant VEGF you are expressing, if the recombinant VEGF from your source does not contain major modifications. The antibody is also guaranteed to work in Western blot.

In summary, I would expect the antibody to work on your protein, although we can not guarantee it.

As an alternative, you might can also look at a polyclonal antibody, which he has several epitopes, such as the ab14078, the ab17696or the ab9570.

https://www.abcam.com/index.html?datasheet=14078 (or use the following: https://www.abcam.com/index.html?datasheet=14078).
https://www.abcam.com/index.html?datasheet=17696 (or use the following: https://www.abcam.com/index.html?datasheet=17696).
https://www.abcam.com/index.html?datasheet=9570 (or use the following: https://www.abcam.com/index.html?datasheet=9570).

I hope this information is helpful. Please do not hesitate to let me know if you have any other questions or concerns.

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Answer

Thank you for your suggestion. I will will put it forward as a suggestion but I think ab1316 would not be the best candidate as having performed sequence analysis of the isoforms this antibody has been shown to recognise (121, 165 and 189) with that of 165b, it seems unlikely that it would be specific for either 165 of 165b. From the file I have attached, The sequence are as follows: P15692-8 isoform 165b P15692-9 isoform 121 P15692-11 isoform 165 P15692-13 isoform 189 From this it can be seen that the 6 amino acid residues of the C-terminal which differentiates between the 165 and 165b isoform are not present at all in isoform 121. I would therefore suspect that the antibody is not binding in this region. I will however have a look through the rest of our catalogue to see if we have any other anti-VEGF antibodies which may be more likely to bind 165 specifically and put these forward for testing. I will let you know of my progress. If you have any further questions or suggestion we would be very pleased to hear them.

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Question
Answer

Thank you for contacting us in regards to Anti-VEGF antibody [VG-1] (ab1316). I’m afraid we currently have no information on whether this antibody would recognise the VEGF isoform 165b. The full length recombinant protein is used as the immunogen and in depth epitope mapping has as yet not been performed. However, as the only difference between the isoforms 165 and 165b is the final 6 C-terminal amino acids I think there is a good chance the antibody recognises a different region. I cannot guarantee it though. We have two antibodies in our catalogue where the VEGF 165b is specifically recognised: ab14994 is a mouse monoclonal which has been used to detect human and rat protein. ab90719 is a rabbit polyclonal which has been used to detect human VEGF 165b. Depending on the applications you intend to use one of these antibodies may be suitable for you. I hope this information has been of help. If you would like any further information please do let me know.

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Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with ab32152 and the order number 984908. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.  

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Answer

Could you confirm that you need ab1316 VEGF antibody; this antibody is not a VEGF receptor 2 specific antibody? I can also suggest trying ethanol as fixative which is far less harsh than acetone; PFA is not a common fixative used in IHC-Fr so you may consider trying ethanol.   I hope this will help.

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1-10 of 24 Q&A

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