I am trying to isolate histones from Hela cells but my preparation is not coming out clean. I am getting other contaminants along with the preparation. Can anyone please come up with suggestion?
too little information to provide you with any suggestion, atleast provide information on the method. Define "not comming out clean" what criteria is used to evaluate this.
are you using total cell extract.
how much histone protein you are trying to isolate
can you isolate the nuclei first then histone
Well by saying , its not coming clean i mean lot of basic proteins are coming along. I am isolating the nuclei first and then acid extracting the histones ..I am aiming at getting a clean preparation where i have only 5 bands of histones. I am straight away taking cells of a confluent T75 flask. I am trying to establish cell culture in my laboratory . So as of now I am not getting into the quantification..
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