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Re: Low phosphate media forBacillus/Bacillus transformation protocol - (Mar/29/2001 )

Hello,It depends which kind of protein you want to extract.I wash the pieces of tissue in PBS ice cold, centrifuge at 4C 3400rpm for 10', then omogenize(very important!) better if with polytron in Suspension Buffer (see Maniatis) or Lysis buffer (no Triton x-100 inside!), boil for 10' and centrifuge at 10,000g for 10'. I use an aliquot of the extract for Bradford assay and then I put the 2x SDS loading buffer (1:1).I hope this could be useful. For every more questions you can write to me.Good Luck!Monica

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does anyone have a general protein extraction protocol for tissue? i have one for cells but not from whole tissue. i want to run the samples on a western. thanks

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