I used to do ERK! Back when half the literature referred to it as MAPK which confused all the microtubule associated protein people.
The fat caterpillar band is exactly what you see when you’ve overloaded the gel. You should drop the load and pull on an appropriate gel to able to see the gap between the 44 and 42 kd species. You may have an effect on just one.
The other issue is that if the lanes have that much protein your controls are all at ceiling. The effect could be due to variable loading but you can’t tell because one lane is at 10x ceiling and the next is at 2x ceiling. The controls look equal but you produce an effect on the target bands. This isn’t even lab meeting quality. This is promising at the end of a shitty day quality.
I used to do ERK! Back when half the literature referred to it as MAPK which confused all the microtubule associated protein people.
The fat caterpillar band is exactly what you see when you’ve overloaded the gel. You should drop the load and pull on an appropriate gel to able to see the gap between the 44 and 42 kd species. You may have an effect on just one.
The other issue is that if the lanes have that much protein your controls are all at ceiling. The effect could be due to variable loading but you can’t tell because one lane is at 10x ceiling and the next is at 2x ceiling. The controls look equal but you produce an effect on the target bands. This isn’t even lab meeting quality. This is promising at the end of a shitty day quality.