cem54

So you're saying that it's likely that I lost most/all the collagen due to a bad purification process? It's entirely possible; it was the first time doing this protocol. And you're suggesting doing something like a BCA before electrophoresis? Ok I will pass that along to my PI. Thank you! Any other suggestions?

Hello all! I am an undergraduate working on a project where we are trying to extract collagen from the fat pad of a human (in the heel). We attempted to extract collagen from fresh tissue samples of a human heel pad using the protocol from Pacak, MacKay, and Cowan (2014, Jove). Then we attempted to electrophoresis the sample in a 7.5% SDS-PAGE gel along with a protein ladder and a sample of standard collagen from Sigma Aldrich (5 mg/mL). When we developed the gel (staining overnight and destaining overnight), all that we saw run was the ladder and the standard from Sigma Aldrich. I am very confused on what might have happened since we followed the procedure exactly and since we also ran samples of the waste that was generated during several parts of the extraction protocol. I am kind of at a loss, so any help would be appreciated. I don't really know what to do next. Thanks!