I am studying a novel protein and we have a probable mwt based on the amino acid seq. However, we have no clue about its glycosylation or phosphorylation level. Neither we know much about its functional role except that its coregulated and coexpressed with TFPI and seems to be playing a role in thrombosis.
Now, we generated 2 primary antibody for it and am getting 2-3 bands in different tissue extracts i.e I have covered almost all tissues/organs (some has 2 some has 3).
I am puzzled up whether all these 2-3 bands are different variants of proteins as its often seen or non-specificity of antibody (we asked a company to make it for us and its affinity purified.....seems to work well in immunostaining)
Question: How can i make sure that the band am looking is the same of my protein?
Is there a way that i remove the glycosylation (if there might be any) and the extra bands might go away in some tissue extracts?