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 4°C 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

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