Can I run GAPDH as a control in western blotting for nuclear protein fraction?
Thank you.
Hi Sajjad, GAPDH is not a nuclear protein. I would use any antibodies directed against histones instead.
good luck, Janine
try histone H3 for nuclear control but it's very strong signal. You can start with 1:10000
Alternatively you could try using lamin B2 or other nuclear structural proteins.
The most common antibodies for nuclear loafing control are histones or laminin b1.
Link for purchase the anti-laminin b1 or Hisyone h3, respectively: http://www.abcam.com/lamin-b1-antibody-nuclear-envelope-marker-ab16048.html
http://www.abcam.com/histone-h3-antibody-nuclear-loading-control-and-chip-grade-ab1791.html
Hi Sajjad,
GAPDH is only good for whole cell lysates and cytoplasmic fractions. You can use histones or laminin b (as suggested by others). You may also consider TBP (TATA binding protein) that I have used myself for nuclear fraction. If you need details of antibody (supplier, dilution etc), please feel free to send a message.
Good luck
We use GAPDH as cytoplasmic control and Laminin A/C as nuclear fraction control in Western blot analysis.
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