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Problem with NE-PER cytoplasmatic and nuclear separation kit - (Apr/05/2011 )


As the topic say i have a problem with this kit (NE-PER Nuclear and Cytoplasmatic separation kit from Thermo scientific). I use this kit on diaphragm and tibialis anterior (leg muscle) from mice. There are no prblems what so ever when I separate the cytoplasm and the nuclei from diaphragm.I get a nice separation in both fractions. I use SOD-1 marker to confirm the purity of cytoplasmic fraction and Histone H1 for the nuclear fraction. But in the TA muscle I cant find none or at least very litle of the Histone HI marker in the nuclear fraction. The cytoplasmatic fraction is about the same as in the diaphragm. So is there anyone else who has experienced the same thing or have experience with this kit? Should i use a different marker? any suggestions on good nuclear markers? are there less H1 proteins in leg muscle? (hard to believe maby)

/ Bettan


Suggestions anyone?