Anti-VEGF Receptor 1 antibody (ab2350)

Thank you for your enquiry. This antibody has not been tested on canine tissues. We do not give discounts on products in order to test the antibody, but we do have a rewards program. If you use the antibody on canine tissue, please consider our AbTrial testing program. For more information see www.abcam.com/abtrial

Thank you for your e-mail. Amplification is a method to amplify signal on immunohistochemical preparations where there is low abundance of the protein of interest (or the primary antibody binding affinity for the protein is low). The most common amplification method is the ABC method, where this procedure employs biotinylated antibody and a preformed Avidin: Biotinylated enzyme Complex and has been termed the “ABC” technique. Because avidin has such an extraordinarily high affinity for biotin (over one million times higher than antibody for most antigens), the binding of avidin to biotin is essentially irreversible. In addition, avidin has four binding sites for biotin, and most proteins including enzymes can be conjugated with several molecules of biotin. The high sensitivity reported for this technique likely resides in either the form or the number of active enzyme molecules associated with the complex and the rapid, irreversible interaction of the complex with biotinylated antibody. In addition to high sensitivity, ABC kits produce very low background staining. In summary, following incubation of the primary antibody (rabbit), wash in PBS your sections/cells, then add an anti-rabbit secondary antibody which is biotin conjugated, incubate for 2-3 hours, then wash in PBS and add HRP-conjugated avidin (provided in a kit, such as the Vector ABC kit). The avidin will bind to the biotin with high affinity and provide an amplification. When DAB will be added at the last step the numerous HRP proteins will give more signal and you will be able to visualise the localisation of your protein of interest.

I would recommend a preliminary experiment using a range of dilutions: 1:200, 1:500, 1:1000, 1:3000 for example and optimise the dilution following the outcome of this experiment.

We do not have any data on the whether it recognises the phospho VEGFR1. We would guess that it would detect both phosphorylated and non-phosphorylated since it was raised against a non-phosphorylated peptide. However, we would be very interested to hear if this is the case

The synthetic peptide sequence for Flt-1 Ab-1: a.a. 1312-1328 (Cys-AELERKIACCSPPPDYN)