Did you run SDS-PAGE for total tissue hogenate? It is good to check the total tissue homogenate, the soluble and insoluble fraction for the presence of proteins using SDS-PAGE. Depending on the physicochemical properties of proteins, some extraction reagents may cause formation of insoluble protein aggregates.
You can check the different extraction reagents for their efficiency in extraction for soluble and stable proteins from tissues and cells.
Thanks Alemu. I used SDS to run the total protein homogenate. This was according to the protocol. Do you have any suggestions to make the protein more soluble ?
formalin crosslinks the proteins in the tumor tissue. It will be very difficult or impossible to extract the proteins with T-PER Tissue Protein Extraction reagent.