Hello, If you fallow Nature protocol for the tube forming assay it is simple and it walk you through the steps you needed for the tube formation assay.

Nature protocols 5:628-635(2010).

I hope it helps.

good luckI

Manju

Hi Kubrah,

You might check the Table-1 from this review paper: Article In Vitro Models of Angiogenesis and Vasculogenesis in Fibrin Gel

In this paper, they summarized fibrin gel-based angiogenic models in Table-1 (fibrinogen and thrombin concentrations, cell types and numbers, and time of the experiment) and you can also find reference numbers there to check protocols in detail. Unfortunately, some of the references cited are related to the bead assay (and I know you are not looking for this) and some are not necessarily to check angiogenesis, but, to develop EC spheroid models (then, of course, these spheroids can be used in angiogenesis assays). However, the protocols from van Hinsbergh (Ref 5) and Greisler (Ref 68) groups might help a bit and can give an idea about fibrin gel-based angiogenesis assays both in 2D and 3D. As an alternative, if you have the chance, you can also order the fibrin-based angiogenesis kit from Merck Millipore: http://www.merckmillipore.com/GB/en/product/Fibrin-In-Vitro-Angiogenesis-Assay,MM_NF-ECM630?ReferrerURL=https%3A%2F%2Fwww.google.co.uk%2F&bd=1

It's nothing special, but, it can give you confidence since there is a well-established protocol and technical support of the company whenever you need. Or, you can use your fibrinogen/thrombin/angiogenic cocktail according to the protocols in the literature and follow company's protocol, as well (I am attaching it). I hope this can help. Good luck!

Thank you very much for your answers. I will try some approaches and will tell about my experiences.