Recombinant human VEGF 165A protein (Active) (ab9571)

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Overview

Description

  • Nature
    Recombinant
  • Source
    Escherichia coli
  • Amino Acid Sequence
    • Accession
      P15692-4
    • Species
      Human
    • Sequence
      APMAEGGGQN HHEVVKFMDV YQRSYCHPIE TLVDIFQEYP DEIEYIFKPS CVPLMRCGGC CNDEGLECVP TEESNITMQI MRIKPHQGQH IGEMSFLQHN KCECRPKKDR ARQENPCGPC SERRKHLFVQ DPQTCKCSCK NTDSRCKARQ LELNERTCRC DKPRR
    • Amino acids
      27 to 191
    • Additional sequence information
      Recombinant human VEGF165 is a 38.2 kDa disulfide-linked homodimeric protein consisting of two 165 amino acid polypeptide chains. It is the full length mature protein minus the signal peptide.

Specifications

Our Abpromise guarantee covers the use of ab9571 in the following tested applications.

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

  • Biological activity

    Determined by the dose-dependent stimulation of the proliferation of human umbilical vein endothelial cells (HUVEC) using a concentration range of 1.0-8.0 ng/ml.

  • Applications

    Cell differentiation

    Cellular Activation

    Functional Studies

    SDS-PAGE

  • Endotoxin level
    < 0.100 Eu/µg
  • Purity
    > 98 % n/a.
    Sterile filtered Greater than 98% pure by SDS-PAGE and HPLC analyses. Endotoxin level is less than 0.1 ng per µg (1EU/µg).
  • Form
    Lyophilised
  • Additional notes
    The biological activity of this product is determined by the dose-dependent stimulation of the proliferation of human umbilical vein endothelial cells (HUVEC) using a concentration range of 1.0-8.0 ng/ml.
  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

    n/a

    This product is an active protein and may elicit a biological response in vivo, handle with caution.

  • Reconstitution
    Reconstitute in 100 µl dH2O

General Info

  • Alternative names
    • glioma-derived endothelial cell mitogen
    • MGC70609
    • MVCD1
    • Vascular Endothelial Growth Factor
    • Vascular endothelial growth factor A
    • Vascular permeability factor
    • VEGF
    • VEGF-A
    • VEGF165
    • Vegfa
    • VEGFA_HUMAN
    • VPF
    see all
  • 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

Images

  • Cell differentiation - Recombinant human VEGF 165A protein (Active) (ab9571)
    Cell differentiation - Recombinant human VEGF 165A protein (Active) (ab9571)This image is courtesy of Sun Yung, Centro de Investigación Príncipe Felipe.

    Human embryonic stem cells (H9) were differentiated both as embryoid bodies and monolayer cultures (on Matrigel) using a protocol described in Kennedy et al., 2006. BMP-4, FGF-basic (ab9596) and VEGF (ab9571) were sequentially added to the media to direct differentiation. These images were taken at day 8, where cell adhesion antigens CD31 (ab32457 in green) and CD34 (ab8536 in red) are abundant and represent primitive haematopoietic cells within the culture. DAPI staining is shown in blue.

References

This product has been referenced in:
  • Jiménez-Torres JA  et al. LumeNEXT: A Practical Method to Pattern Luminal Structures in ECM Gels. Adv Healthc Mater 5:198-204 (2016). Read more (PubMed: 26610188) »
  • Tan J  et al. Extracts of endophytic fungus xkc-s03 from Prunella vulgaris L. spica inhibit gastric cancer in vitro and in vivo. Oncol Lett 9:945-949 (2015). Read more (PubMed: 25624914) »
See all 4 Publications for this product

Publishing research using ab9571? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-8 of 8 Abreviews or Q&A

Question

I have a scaffold that contains sulfated glycoaminoglycans (GAGs). GAGs can bind VEGF in vivo. I need to absorb VEGF to these scaffolds so the first question is if VEGF ab9571 binds to GAGs in its form (Is it a dimer? Does it have the GAG attachment cites etc.).
After, I need to immuno-stain the VEGF with the proper antibody ab 1316 and take pictures using fluorescent microscope. So the second question is whether ab1316 will stain VEGF ab9571.
After, I would like to stimulate HUVEC with VEGF ab9571. So the third question is whether this form of VEGF ab9571 will properly stimulate these cells.

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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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Question

Dear All,



Are those proteins below injectable in Mice?




1. SPARC protein (MBP) (ab92965)



2. TGF beta 1 protein (Active) (ab50036)




3. VEGF protein (Active) (ab9571)

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Answer

Thank you for contacting us and for your patience.

I had to contact the laboratory in order to get more information about the use of these three proteins.

I can confirm, that for the ab92965 SPARC protein (MBP) to our knowledge it has not been tested for injection in mice or other animals (besides it uses as immunogen for polyclonal antibodies production in rabbits intramuscular). It could be that it will work for injection in mice but we can not guarantee this and we have no information about different administration routes (intravenous, intramuscular etc.).

The two other proteins have been used in mice before. See the following links:

1. ab9571 VEGF protein (Active)

http://www.ncbi.nlm.nih.gov/pubmed/19088027

http://www.ncbi.nlm.nih.gov/pubmed/10784613

http://www.ncbi.nlm.nih.gov/pubmed/11689883

http://www.ncbi.nlm.nih.gov/pubmed/12036930

http://www.ncbi.nlm.nih.gov/pubmed/9401980

http://www.ncbi.nlm.nih.gov/pubmed/15358628

http://www.ncbi.nlm.nih.gov/pubmed/16477043

http://www.ncbi.nlm.nih.gov/pubmed/16888632

http://www.pnas.org/content/104/8/2837.full

http://www.ncbi.nlm.nih.gov/pubmed/18180379

http://www.ncbi.nlm.nih.gov/pubmed/15067321

http://www.ncbi.nlm.nih.gov/pubmed/20056792

http://www.ncbi.nlm.nih.gov/pubmed/22025303

http://www.pnas.org/content/early/2011/07/25/1016546108.abstract



2. ab50036 TGF beta 1 protein (Active)

http://www.ncbi.nlm.nih.gov/pubmed/20304823

http://www.ncbi.nlm.nih.gov/pubmed/22761317

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

Can you please send me the sequences for these VEGF proteins?

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Answer

Thank you for contacting us. The sequences are as follows:
ab9571: two polypeptide chains with the sequence APMAEGGGQN HHEVVKFMDV YQRSYCHPIE TLVDIFQEYP DEIEYI FKPS CVPLMRCGGCCNDEGLECVP TEESNITMQI MRI KPHQGQH IGEMSFLQHN KCECRPKKDR ARQENPCGPC SERRKHLFVQ DPQTCKCSCK NTDSRCKARQ LELNERT CRC DKPRR
ab61861: APMAEGGGQN HHEVVKFMDV YQRSYCHPIE TLVDIFQEYP DEIEYIFKPS CVPLMRCGGC CNDEGLECVP TEESNITMQI MRIKPHQGQH IGEMSFLQHN KCECRPKKDR ARQENPCGPC SERRKHLFVQ DPQTCKCSCK NTDSRCKARQ LELNERTCRC DKPRR
ab62134: double chain with the sequence MAPTTEGEQK SHEVIKFMDV YQRSYCRPIE TLVDIFQEYP DEIEYIFKPS CVPLMRCAGC CNDEALECVP TSESNITMQI MRIKPHQSQH IGEMSFLQHS RCECRPKKDR TKPEKHCEPC SERRKHLFVQ DPQTCKCSCK NTDSRCKARQ LELNERTCRC DKPRR
ab83572: APMAEGGGQN HHEVVKFMDV YQRSYCHPI ETLVDIFQEY PDEIEYIFKP SCVPLMRCG GCCNDEGLE CVPTEESNIT MQIMRIKPHQ GQHIGEMSF LQHNKCECRP KKDRARQEN PCGPCSERRK HLFVQDPQTC KCSCKNTDSR CKARQLELNE RTCRCDKPRR
ab89574: APMA EGGGQNHHEV VKFMDVYQRS YCHPIETLVD IFQEYPDEIE YIFKPSCVPL MRCGGCCNDE GLECVPTEES NITMQIMRIK PHQGQHIGEM SFLQHNKCEC RPKKDRARQE NPCGPCSERR KHLFVQDPQT CKCSCKNTDS RCKARQLELN ERTCRCDKPR R

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Question

Dear technical team,

Our customer would like to purchase VEGF protein and he has CST #8065.

http://www.cellsignal.com/products/8065.html

I searched alternative item ab9571. It is also 165 amino acids but there's not detail information so,let me know the sequence information.

CST : Recombinant human VEGF165 (hVEGF165) Ala207-Arg371 (Accession #NP_001020539) was expressed in human 293 cells at Cell Signaling Technology.

Best regards,

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Answer

Thank you for contacting us and your interest in our products.

We have a few different products which may be of interest to your customer. Proteins ab61861, ab56620 and ab83572 are all of the VEGF 165A form. Additionally, ab61861 and ab83572 have both been shown to have biological activity as measured by using a HUVEC assay. I think these two may therefore be of most interest to your customer.

I hope this information has been of help. If you require any further information, please do not hesitate to contact us again.

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Question

I am interested in VEGF165 (recombinant expressed in E. coli) as well as in an Anti-VEGF-antibody. I would like to do conformation analysis and I need the mature VEGF-protein with length of 165 amino acids. So there should be no N-terminal signal sequence.

You offer these three VEGF165 proteins: ab56620, ab61861, ab9571 and I wonder which of your products is the right one for me. Do ab56620 and ab61861 contain 165 or 191 aa?

Further which antibody (preferred host: rabbit) do you recommend to detect the mentioned proteins?

Can you give me any further sequence information about the target proteins of following antibodies: ab52917, ab9570, ab99511?

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Answer

Thank you for contacting us.

For the proteins, I would suggest using ab61861 as this protein is arecombinant Human VEGF 165A protein (double, non-glycosylated, polypeptide chain containing 165 amino acids, MW 38kDa, expressed in E.coli).

In terms of the most suitable antibodies, ab9570 which is reactive with VEGF 165and 121, would be the best choice. It was raised against highly pure (>98%) recombinant hVEGF 165 (human Vascular Endothelial Cell Growth Factor), however, the epitope has not been mapped.

As for ab52917 and ab99511, I willcontact the lab to obtain more information about the immunogen region. As soon as I have more details, I will let you know.

What kind of conformation studies are you planning to do? As far as I can tell such application has not been tried with neither these protein nor antibodies.

I hope this information is already of some help to you.
In the meantime, please do not hesitate to contact us if you need any more advice or information.

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Question

Need a antibody a protein match pair for VEGF

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Answer

Thank you for contacting us today. I can confirm that the VEGF protein ab9571 has been used in ELISA with the VEGF antibody ab9570. Therefore they are a suitable match and ab9571 can be used to generate a standard curve for ELISA. Please let me know if you need further information.

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Question

I have a new antibody for pVEGFR2. I wanted to grow HUVEC cells as a positive control for this antibody. To do so I believe I need to add VEGF to stimulate growth. If I added this VEGF protein to HUVEC cells in culture an I correct in thinki8ng that it would signal through VEGFR2 and thus would phosphorylate this receptor tyrosine kinase?

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Answer

Thank you for your enquiry. Please see the response from the originator of the recombinant protein: Regarding our human VEGF, we test this by the dose-dependent stimulation of the proliferation of human umbilical vein endothelial cells (HUVEC) using a concentration range of 1.0-8.0 ng/ml. We have not tested for phosphorylation. Please find a link below with reference to using our recombinant human VEGF activating the VEGFR2: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=14764537&dopt=Abstract I hope this information helps. If you have any further questions please let me know.

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Question

How should this protein be reconstituted? How pure is it?

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Answer

Thank you very much for your enquiry and patience. Ab9571 should be reconstituted with water to a concentration of 0.1-1.0 mg/ml. Also, this product was sterile filtered and it is greater than 98% pure by SDS-PAGE and HPLC analyses. Please contact us again if you have any additional questions.

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