Protocol Online logo
Top : New Forum Archives (2009-): : SDS-PAGE and Western Blotting

Low Molecular Weight Housekeeping Gene for Loading Control? - (Oct/19/2011 )

Hello all,

I am wondering if anyone knows a good housekeeping gene to use in western blotting from whole cell lysates that is a very low molecular weight? I cannot use actin, as I normally would, because my protein of interest is 45 kD. Preferably, (since I ran out the dye and the lowest band from my ladder is 20 kD) I am interested in something between 20 and 25 kD, give or take a few kD.

Thank you in advance,


I should add that my gel has a number of (as of yet) unspecified bands, one of which shows up around 37 kD, so I'm not sure that even GAPDH would work.


Tubulin has a band of aprox 50 KDa, maybe you can use it, or GAPDH if you change the secondary antibody (if your first Ab is made in rabbit, you can buy a GAPDH Ab in mouse)
Hope it helps you

-Irene G-