I'm performing standard histone extraction (abcam protocol) and western blot essay on my HepG2 cells but the band's that I get don't seem to have the classic histone shape.
Any clue guys?
Hi Guglielmo,
Shape or molecular weight? Which were the sample treatments and SDS-PAGE conditions? Is the image a Ponceau S staining? Have you visited the https://www.abcam.com/protocols/western-blot-faq page to analyze possible factors involved?
How much cells did you use? This looks pretty small amount of histones. What sizes are your bands at? ~15kDa?
I assume you are doing acid extraction. Did you neutralize the acids completely? Did you then boil samples in SDS/PAGE buffer to denature? Also, Ponceau S is NOT sensitive; try staining with comassie blue and you will see many more bands. Lastly, is your lowest marker 20kD, if so you may have run some histones off the gel.
- Badges
- Science method