I want to extract an extracellular protein from tissue to be further subjected to gel filtration and subsequently mass spec in other to identify potential interaction partner.
As this protein is mostly expressed during development , there seems to be enough tissue material to extract enough protein for my experiments and moreover it seems this protein is highly insoluble and cannot be easily extracted.
What approach can I use to enrich the amount of protein needed for my subsequent experiment.
Thanks
Which tissue are you going to use to extract the proteins? Protocol used for one specific tissue type may not work for other tissue type because of different ECM complexities and other factors. I guess there are lots of protocol on online, you can get the protocol of your choice to extract the proteins from your tissue of interest.
I am looking at different tissue ranging from skin, kidney, new born head etc.
It will be appreciated if you can share a link of useful protocol.
What ECM protein?? There are between 50-100 of them and most require a specific isolation process.
ECM proteins in "large quantity" are mostly collagens, fibronectin, laminins, elastin and proteoglycans.
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