I work a lot with histones, and I can't seem to get consistent western blot results with the same sample of histones. I've ran these samples on a gel 3 times and got different results every time. The samples are about a month old now.
I did W.B for histone proteins from cells/ cell lysate that kept in -80 for more than 6 months and they are very stable. For more information, check my following paper.
Article Systematic Proteomic Identification of the Heat Shock Protei...
You did not mention how you stored samples during whole month, RT, 4C or -20 0r -80C Or how many freeze/thaw cycle etc.
I agree, you need to store your histone samples in -20 or better in -80 freezer for long term storage. But I think histones in SDS sample buffer stored in -20 freezer should be OK for many months. I am not quite sure how you stored your samples and what your gel running and transfer conditions were? But if you kept everything consistent, you should be able to get consistent results. I would also like to say that when you re-use your SDS samples stored at -20, you need to warm up the samples so that everything re-dissolves into solution and mix well before you load the samples on your gel. Good luck with your experiments!
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