Recombinant human VEGF Receptor 2 protein (Active) (ab106310)

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.

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.

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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.