I am having trouble getting a detection with my western blotting.
I am looking for around 70kda.
I have gone over it a million times and just can not get a good signal except once and when I did it was with a new blocking solution but after that I could not get a signal.
I use 10% gel with a bio rad apparatus, to transfer I keep everything cold I use 50mL of 10 transfer buffer, 200mL methanol, and 750 mL water.
I use .45 nitro membrane at 100V for 1 hour.
To detect ECL kit.???