Overview

  • Product name
    Anti-VEGF Receptor 2 antibody
    See all VEGF Receptor 2 primary antibodies
  • Description
    Rabbit polyclonal to VEGF Receptor 2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, IHC-P, IHC-Fr, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Dog, Human
  • Immunogen

    Synthetic peptide:

    KMVDAAVHADSGTTLQLTSC

    , corresponding to C terminal amino acids 1326-1345 of Mouse VEGF Receptor 2.

  • Positive control
    • Angiosarcoma.
  • General notes

    Isoform 1 localization: Cell membrane; Single-pass type I membrane protein. Cytoplasm. Nucleus. Cytoplasmic vesicle. Early endosome.

    Note: Detected on caveolae-enriched lipid rafts at the cell surface. Is recycled from the plasma membrane to endosomes and back again. Phosphorylation triggered by VEGFA binding promotes internalization and subsequent degradation. VEGFA binding triggers internalization and translocation to the nucleus.

Applications

Our Abpromise guarantee covers the use of ab2349 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
IP 1/100.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr 1/50 - 1/100.
ICC/IF Use at an assay dependent concentration. See Abreview.
Flow Cyt Use at an assay dependent concentration. PubMed: 18602918

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Receptor for VEGF or VEGFC. Has a tyrosine-protein kinase activity. The VEGF-kinase ligand/receptor signaling system plays a key role in vascular development and regulation of vascular permeability. In case of HIV-1 infection, the interaction with extracellular viral Tat protein seems to enhance angiogenesis in Kaposi's sarcoma lesions.
  • Involvement in disease
    Defects in KDR are associated with susceptibility to hemangioma capillary infantile (HCI) [MIM:602089]. HCI are benign, highly proliferative lesions involving aberrant localized growth of capillary endothelium. They are the most common tumor of infancy, occurring in up to 10% of all births. Hemangiomas tend to appear shortly after birth and show rapid neonatal growth for up to 12 months characterized by endothelial hypercellularity and increased numbers of mast cells. This phase is followed by slow involution at a rate of about 10% per year and replacement by fibrofatty stroma.
  • Sequence similarities
    Belongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily.
    Contains 7 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    Phosphorylated. Dephosphorylated by PTPRB. Dephosphorylated by PTPRJ at Tyr-951, Tyr-996, Tyr-1054, Tyr-1059, Tyr-1175 and Tyr-1214.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD309 antibody
    • CD309 antigen antibody
    • EC 2.7.10.1 antibody
    • Fetal liver kinase 1 antibody
    • FLK-1 antibody
    • FLK1 antibody
    • FLK1, mouse, homolog of antibody
    • Kdr antibody
    • Kinase insert domain receptor (a type III receptor tyrosine kinase) antibody
    • Kinase insert domain receptor antibody
    • KRD1 antibody
    • Ly73 antibody
    • Protein tyrosine kinase receptor FLK1 antibody
    • Protein-tyrosine kinase receptor flk-1 antibody
    • soluble VEGFR2 antibody
    • Tyrosine kinase growth factor receptor antibody
    • Vascular endothelial growth factor receptor 2 antibody
    • VEGFR 2 antibody
    • VEGFR antibody
    • VEGFR-2 antibody
    • VEGFR2 antibody
    • VGFR2_HUMAN antibody
    see all

Images

  • ab2349 staining the VEGF Receptor 2 in Mouse skin tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (1/100 in PBS + 2% blocking serum) for 16 hours at 4°C. A biotin-conjugated Goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence analysis of human cardiomyocytes labeling VEGF Receptor 2 with ab2349 at 1/400 dilution. Cells were fixed with acetone and blocked with 5% serum for 1 hour at 25°C. Cells were incubated with ab2349 at 1/400 for 24 hours at 4°C in 10% FBS. A polyclonal goat anti-rabbit IgG Alexa Fluor® 488 conjugate was used as the secondary antibody at 1/1000 dilution.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human normal breast tissue labeling VEGF Receptor 2 with ab2349 at 1/200 dilution. Sections were lightly counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human normal breast tissue labeling VEGF Receptor 2 with ab2349 at 1/200 dilution. Sections were lightly counterstained with hematoxylin.

  • ab2349 at a 1/100 dilution staining Human choroidal endothelial cells by Immunocytochemistry. The antibody was incubated with the cells for 3 hours and then detected using an Alexa-Fluor 488® goat anti-rabbit (IgG).

    This image is courtesy of an Abreview submitted on 27 February 2006.

    See Abreview

References

This product has been referenced in:
  • Hinderer S  et al. Surface functionalization of electrospun scaffolds using recombinant human decorin attracts circulating endothelial progenitor cells. Sci Rep 8:110 (2018). Read more (PubMed: 29311692) »
  • Yu B  et al. CCDC134 serves a crucial role in embryonic development. Int J Mol Med 41:381-390 (2018). Read more (PubMed: 29115376) »
See all 66 Publications for this product

Customer reviews and Q&As

1-10 of 29 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (First trimester human placenta)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate
Specification
First trimester human placenta
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 29 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (Granulation tissue)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
Granulation tissue
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 30% · Temperature: 25°C
Fixative
Formaldehyde

Francisco Fernandez Flores

Verified customer

Submitted Dec 01 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (iCell® Cardiomyocytes)
Permeabilization
No
Specification
iCell® Cardiomyocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Nov 25 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Antigen retrieval step
None
Sample
Rat Tissue sections (skin)
Specification
skin
Permeabilization
No
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jun 10 2014

Question

Dear Techserve, kindly receive customers reply below:

Thank You for Your reply! Please look below for the answers.

1. Has the protein transfer to the membrane and quality of the sample been assessed using a loading control?

We always use a loading control for western blot. The pdf document that I had sent to you earlier indicates beta actin as a loading control clearly.

2. Is the current vial of secondary antibody working well with other primary antibodies ?

Yes, it does. We are running several western blottings every day, so our secondary antibodies are almost daily tested by several colleagues.

The host species for the abcam c-MET primary antibody is goat. For beta actin primary antibody , the host species is goat as well. The secondary antibody used was the same for both these primary antibodies (donkey anti-goat). The beta actin bands are very clear, whereas there were no bands for c-MET. This clearly indicates that the current secondary antibody vial works.

Similarly, the rabbit secondary antibody used for flow cytometry has been in continuous use in our lab and the current vial works.

3. Increasing the antibody concentrations

Regarding the western blotting: if the signal is very mild or weak, then we can enhance the signal using a higher primary antibody concentration, but if we have no signal (even with the femto chemiluminiscence detection), then I personally have doubt that increasing the antibody concentration will help. With respect to flow cytometry, increasing the concentration of primary antibody, we have to increase the corresponding IgG control, which in turn may result in proportional higher signal. Nevertheless, I will take your suggestion and use a higher concentration of the antibodies.

In the meanwhile, could we kindly proceed with this decision of replacement of the antibodies.

Await your reply thankfully!

Read More
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 the products 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 the order number ######.

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.

Read More

Question
Answer

Thank you for taking the time to complete our questionnaire.

The details you have kindly provided will provide us with vital information for our monitoring of product quality.

I appreciate the time the customer have spent in the laboratory and understand their concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there very few tips to provide on this occasion to help improve the results from either of these antibodies. I can suggest it is possible you may have regrettably received a bad vial on this occasion.

However, I can recommend it would be important to consider the following:

ab10728 WB
1. Has the protein transfer to the membrane and quality of the sample been assessed using a loading control?
2. Is the current vial of secondary antibody working well with other primary antibodies?
3. I can suggest to try increasing the antibody concentration, for example 0.5 or even 1 ug/ml. The dilution provided on the datasheet is a guideline only. Increasing the concentration may help increase the signal.

ab2349
1. Antibodies are usually used at quite a high concentration in flow cytometry, often at a dilution between 1:10 to 1:50. Has this been tried? It may increase the signal obtained.
2. Is the current vial of secondary antibody working well with other primary antibodies?

Alternatively, or if these tips do not work, I am pleased to offer you a free of charge replacement or credit note for each antibody in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that our antibody are tested and covered by our 6 month guarantee for the tested applications and species listed on the individual datasheets. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I appreciate your concerns and it is regrettable the results have not been successful. Before deciding how to proceed in this case, I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.

WESTERN BLOT:


Order Details
Antibody code:

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


Amount of protein loaded


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.




FLOW CYTOMETRY

Order Details
Antibody code:


Problem:
Choose: Non-specific staining No signal or weak signal High background


Lot number

Purchase order number
or preferably Abcam order number


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, low signal, no signal etc.)


Sample (Species/Cell type/Cell line etc.)


Sample preparation for single cell preparation (Buffer etc.)


Number of cells used


Permeabilization, fixation step


Blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method


Positive and negative controls used (please specify)




Optimization attempts (problem solving)
How many times have you tried the FACS?



Have you run a "No Primary" control?


Yes No
Do you obtain the same results every time?
Yes No

What steps have you altered?


Additional Notes:


Data:
We would appreciate if you are able to provide any data/histograms which will help us to assess the results

Read More

Answer

Thank you for your message and for providing this further information.

I appreciate the time your customerhas spent on these experiments and would be pleased to arrange in compensationa credit note ora replacement with either a different lot of ab2349 or an alternative anti-VEGF receptor 2 such as ab39256, ab39638, ab131441, ab45010 orab131510.

I look forward to hearing from you with details of how you and your customerwould like to proceed.

Read More

Answer

Thank you for contacting Abcam regarding ab2349.

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.

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.

Read More

Answer

Thank you for your reply. xxxxis currently away from the office but I hope I can provide you with the information you have requested.

The Anti-VEGF Receptor 2 antibody (ab2349) has only so far been validated inhouse with human tissue in IHC. The information regarding its reactivity with rat samples has come from customer feedback as well as published articles:

Mafuvadze B et al. Apigenin prevents development of medroxyprogesterone acetate-accelerated 7,12-dimethylbenz(a)anthracene-induced mammary tumors in Sprague-Dawley rats. Cancer Prev Res (Phila) 4:1316-24 (2011). PubMed: 21505181

Benakanakere I et al. Synthetic Progestins Differentially Promote or Prevent 7,12-Dimethylbenz(a)anthracene-Induced Mammary Tumors in Sprague-Dawley Rats. Cancer Prev Res (Phila Pa) 3:1157-67 (2010). PubMed: 20699413

Seabrook TJ et al. Angiogenesis is present in experimental autoimmune encephalomyelitis and pro-angiogenic factors are increased in multiple sclerosis lesions. J Neuroinflammation 7:95 (2010). PubMed: 21176212

Koleganova N et al. A calcimimetic (R-568), but not calcitriol, prevents vascular remodeling in uremia. Kidney Int 75:60-71 (2009). PubMed: 19092814

Cheluvappa R et al. The effect of old age on liver oxygenation and the hepatic expression of VEGF and VEGFR2. Exp Gerontol 42:1012-9 (2007). PubMed: 17658234

The IHC-P protocol followed with this antibody inhouse was performed using HIER withCitrate Buffer pH 6.0 and the antibody was detected using an HRP polymer using DAB and hematoxylin used as a counter stain.

If you require any further information please do let us know.

Until then, I wish you all the best with your research.

Read More

1-10 of 29 Abreviews or Q&A

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

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