What is a suitable nuclear/ cytoplasmic loading control to use for westerns on nuclear lysates made from raw cells for detecting nfkb activation and its subunit expression?
we have also used nucleoporin 62 on immunoblots when assessing sub cellular fractionation.
Have you considered to use the total protein of your sample as loading control to avoid the known problems with the so called "house keeping proteins"? If you have the possibility to image fluorescence in your lab, you could try to label your total protein (in a minimal labeling, so there is no interference with your antibody binding) to co-detect it together with your target. This gives you a statistically valid normalization compared to using a single protein like actin, of which you you can't be sure if it is regulated or not. If you want to know more have a look on the following link:
http://www.dyeagnostics.com/site/en/products/spl/
Check out the following link, there's a nice table showing loading controls for various cell fractions:
https://www.linkedin.com/company/novatein-biosciences-inc
- Badges
- Science method