Overview

Description

  • Nature
    Recombinant
  • Source
    Insect cells
  • Amino Acid Sequence
    • Accession
    • Species
      Human
    • Sequence
      ASVGLPSVSLDLPRLSIQKDILTIKANTTLQITCRGQRDLDWLWPNNQSG SEQRVEVTECSDGLFCKTLTIPKVIGNDTGAYKCFYRETDLASVIYVYVQ DYRSPFIASVSDQHGVVYITENKNKTVVIPCLGSISNLNVSLCARYPEKR FVPDGNRISWDSKKGFTIPSYMISYAGMVFCEAKINDESYQSIMYIVVVV GYRIYDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQH KKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNS TFVRVHEKPFVAFGSGMESLVEATVGERVRIPAKYLGYPPPEIKWYKNGI PLESNHTIKAGHVLTIMEVSERDTGNYTVILTNPISKEKQSHVVSLVVYV PPQIGEKSLISPVDSYQYGTTQTLTCTVYAIPPPHHIHWYWQLEEECANE PSQAVSVTNPYPCEEWRSVEDFQGGNKIEVNKNQFALIEGKNKTVSTLVI QAANVSALYKCEAVNKVGRGERVISFHVTRGPEITLQPDMQPTEQESVSL WCTADRSTFENLTWYKLGPQPLPIHVGELPTPVCKNLDTLWKLNATMFSN STNDILIMELKNASLQDQGDYVCLAQDRKTKKRHCVVRQLTVLERVAPTI TGNLENQTTSIGESIEVSCTASGNPPPQIMWFKDNETLVEDSGIVLKDGN RNLTIRRVRKEDEGLYTCQACSVLGCAKVEAFFIIEGA
    • Molecular weight
      116 kDa
    • Amino acids
      20 to 757

Specifications

Our Abpromise guarantee covers the use of ab54347 in the following tested applications.

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

  • Applications

    SDS-PAGE

    Functional Studies

  • Purity
    > 95 % SDS-PAGE.
    ab54347 was visualised on SDS Page by Silver stain.
  • Form
    Lyophilised
  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

    pH: 8.00
    Constituents: 0.121% Tris, 0.58% Sodium chloride

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

  • Reconstitution
    Reconstitute in water or PBS to a concentration of not lower than 100 µg/ml.

General Info

  • Alternative names
    • CD309
    • CD309 antigen
    • EC 2.7.10.1
    • Fetal liver kinase 1
    • FLK-1
    • FLK1
    • FLK1, mouse, homolog of
    • Kdr
    • Kinase insert domain receptor
    • Kinase insert domain receptor (a type III receptor tyrosine kinase)
    • KRD1
    • Ly73
    • Protein tyrosine kinase receptor FLK1
    • Protein-tyrosine kinase receptor flk-1
    • soluble VEGFR2
    • Tyrosine kinase growth factor receptor
    • Vascular endothelial growth factor receptor 2
    • VEGFR
    • VEGFR 2
    • VEGFR-2
    • VEGFR2
    • VGFR2_HUMAN
    see all
  • 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

Images

References

ab54347 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Question

Dear Tech support

Attached is a customer complaint form for ab106041.
Here is what the customer has written in addition:



I tested the VEGF receptors (ab54346, ab106310 & ab54347) with the Rabbit anti-VEGF-HRP (ab106041) in ELISA that contain: receptors coated the plate with 0.5-1µg/ml followed by BSA blocking and VEGF165 2-0.002µg/ml . Detection (Rabbit anti-VEGF-HRP) tested 1:5,000 & 1:10,000. I get no signal at all in all wells.

Please specify as many details as possible:



Order Details

Antibody code: ab106041



Problem (Problem with standard curve/High Back ground/No Signal or weak signal):

Batch number: GR86046-2

General Information

Antibody storage conditions (temperature/reconstitution etc): -20

Description of the problem (high background, no signal, non-specific color development, poor standard curve etc.):

Type of ELISA (Direct ELISA/Indirect ELISA/Sandwich ELISA etc.)

Sample (Species/Cell type/Cell line etc.): VEGF 2-0.002µg/ml

Coating well (Buffer/Concentration of the coating material etc.): VEGF receptor 1 (Cat# ab54346 Lot#GR48382-1), VEGF receptor 2 (Cat# ab10631 Lot#GR52382-4) & VEGF receptor 2 Active (Cat# ab54347 Lot# GR24004-3).

0.5 & 1µg/ml ; 100µl/well for overnight at 2-8°C.

Blocking conditions (Buffer/time period, Blocking agent etc.): 1% BSA in 0.05%tween in PBS ; 200µl for 2 hours at 37°C with 650RPM shaking.

Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step): ab106041 Rabbit anti-VEGF – HRP 1:5000 & 1:10,000 ;100µl for 1 hour at 37°C with 650RPM shaking

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

Detection method (Substrate/Diluent etc.) 100µl/well TMB

Positive and negative controls used (please specify):

Optimization attempts (problem solving)

How many times have you tried the ELISA? twice

Have you run a "No Primary" control?

Do you obtain the same results every time? yes

What steps have you altered?

Additional Notes:

Could you have an explanation for this?

How does abcam tested these products except antibody against the receptors?
Please advise,
Regards,

Read More
Answer

Thank you for your enquiry regarding ab106041 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

After reading through the detailed protocol you kindly forwarded to Abcam, I would like to make the following comments/suggestions:

1) The experimental set up is not the standard Sandwich ELISA, it looks like an activity assay. Since in the sELISA, a pair of primary antibodies (capturing and detection) are normally used to capture the antigen. In this case there is only one primary antibody and the antigen which is coated directly to the wells is not VEGF - but the receptor 1 and 2.

2) ab106041 is Rabbit polyclonal to VEGF (HRP) and since the immunogne was a recombinant full length Human VEGF protein will recognize both native and recombinant Human VEGF (not the receptor protein) but not the receptor proteins.

3) Tested application:

ab54346 - SDS-PAGE only,

ab54347- SDS-PAGE only,

ab106310- functional assay and SDS-PAGE (not ELISA)

Q1: Has the customer coated the ELISA plates with human VEGF protein?

It would be worth checking if the HRP enzyme is still active by adding substrate directly to a few ul of the ab106041

I hope this helps and if I can assist further, please do not hesitate to contact me.

Read More

Answer

Thank you for contacting us.

The antibodies are viable for 10 years when stored at -20C, for 1 year when stored at 4C.

Please note that we recommend storing the antibodies at 4C for 1-2 weeks only. Some antibodies are prone to degradation if stored longer than a month so please check the datasheet for storage conditions.

ab106041; Store at -20°C, Aliquot to avoid repeated freezing and thawing. will be viable for 10 years
ab106310; Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. will be viable for 10 years at -20 or -80C.
ab51516; Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles; viable for 10 years
ab54346; Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles; viable for 10 years at -20C.
ab54347; Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles; viable for 10 years at -20C
ab83132; Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles; viable for 10 years at -20C

Click the link for more up-to-date information:

https://www.abcam.com/index.html?pageconfig=resource&rid=10795

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Question
Answer

Thank you for contacting Abcam regarding this.

Unfortunately because this information is considered proprietary, I am not able to share the exact contents of the standard/sample diluent other than to tell you it is an animal serum with 0.09%sodium azide as preservative. We also do not currently carry the peptide used as standard for individual purchase.

We do carry a VEGF Receptor 2 protein (Active) (ab54347) thatconsists of all 7 extracellular domains, which contain all the information necessary for high affinity ligand binding. When developing an assay of your own, I would highly recommend this product as a standard. The product page for this is:https://www.abcam.com/VEGF-Receptor-2-protein-ab54347.html

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Use our products? Submit an Abreview. Earn rewards!

https://www.abcam.com/abreviews

Read More

Answer

Thank you for your patience while I looked into your question. I have heard back from the lab, and ab106310 is also only the extracellular domain of VEGF receptor 2, and is thus not suitable for kinase studies. However, ab42614 has been tested for kinase activity and would be a good choice for your studies. The phosphorylation activity has been determined as 40 U/µg, where one unit is defined as the amount of enzyme that will transfer 1 pmol phosphate to Tyr substrate per minute at pH 7.4 and 30°C. I hope this information is helpful, but please let me know if you have any questions.

Read More

Answer

Thank you for contacting us with your question. Ab54347 is composed of the 7 extracellular ligand binding domains, and it does not contain the intracellular tyrosine kinase domain. So, this protein would not be suitable for kinase assays. We have another VEGF receptor 2 protein, ab106310, and I have emailed the production laboratory to inquire whether this protein would be suitable for kinase assays. I will let you know their response as soon as I hear back from them. Let me know if you have any questions.

Read More

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

Sign up