Nature of total proteins can influence the migration? - (Mar/05/2007 )
I made a western blot with 2 differents types of proteins extracted from 2 différents species (human heart tissus and hamster cultured cell line). (same denaturation with same buffer, good dosage)
Is there some reason why my proteins don't migrate at the same rate? (hamster proteins a bit faster than human)
Does the nature of total proteins could influence the rate of migration?
I made 2 gels and I had the same results
Different species could modify protiens such that they might run differently. Also if one well has more protein than other, this could also influence the way they run.
You have used total protien extact from a tissue and compared it to protien extract from cell line. Tissue would contain much more protien and could have modified your protien which might not be in the case of the cell lines.
Could you find any publication about it? It would be very useful for me!
For now, i don't find anything....
scolix is right, if you load more proteins, your protein migrates less. (and separation is less good)
I don't have a paper, I only know that by personal experience.
Yes I know it
But my interrogations was about : " Different species could modify protiens such that they might run differently".
If I consider my dosage right, why It ran differently?
If I recall correctly my biochemistry, proteins in both species should be charged equally with SDS. So, if you load an equal amount of proteins in both wells, same weight proteins should migrate at the same rate. May the hamster protein be post-traductionally modified by a protease and be shorter than it is in human?
I dont have any papers. Well, proteins in human are different from mouse (not all protiens but some and these have different sequence, which could also lead to specific post translational modification). And their behavior could also be different. Eg. Human a-synuclein can aggregate quite easily as compared to mice.