What is the best loading control can be used proteins in membrane fraction other than catenin proteins
Earlier we did subcellular fractionation by differential centrifugation. That literature has good membrane markers-please check it. It will help you distinguish plasma membrane marker from mitochondrial membrane marker, ER marker etc...Hope this helps
You could use ab against integrin or cadherin subunits. Just be carefull when selecting antibody because both have cytoplasmis domains
Thank you every one for your kind replies!! But I'm looking at Adherence junction difference in my knockout mice, thus I cannot use any of the cadherens nor catenins. Again Na+/K+-ATPase run at the same molecular weight as most if the catenin and cadherin markers I want to look at, so I won't be able to use that either. So any more suggestion guys (other than transfer in)??
Have you tried one of the standard markers like beta-actin? I'm pretty sure that cytoskeleton would likely come down with membrane fractions. Is it really an issue if the markers suggested run at the same molecular weight as your other markers? All you have to do is strip the western blot and reprobe with your loading control gene - if the antibodies are good enough, it will not be an issue.
I agree with Dr. Clarke. The cytoskeleton with pull out beta actin like soil clinging to roots of a plant. Try to use primary antibodies from different species. This will allow you to take advantage of a dual color system such as a Licor Odyssey or Bio-Rad Versa Doc.
Good luck,
Chris
Earlier we did subcellular fractionation by differential centrifugation. That literature has good membrane markers-please check it. It will help you distinguish plasma membrane marker from mitochondrial membrane marker, ER marker etc...Hope this helps
Hey Venil thanks for ur kind reply! Can u please send me the reference of that paper please. Thank you !
Maybe this will help?
http://www.abcam.com/primary-antibodies/loading-control-guide
You can use NaK ATPase. Voltage-dependent anion channels (VDAC 1, VDAC 2, VDAC 3 may also be used. APX3 (ascorbate peroxidase 3) has been used as a loading control for plant membrane fractions.
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