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==Materials==
*RIPA Buffer (see [[Buffer/RIPA|RIPA]])
*Mouse Tissues
==Protocol==
#Weigh frozen tissue samples, only need 100-300 mg of tissue. If there is too much cut it off and return the extra tissue to the -80
#For fibrous tissues chop into small pieces with scissors
#Add 3 volumes of RIPA to tissue and homogenize 20x with 1 mL glass homogenizer (normally on SHC bench)
#Transfer lysate to fresh tube and keep on ice
#Clean homogenizer and lyse remaining tissues
#Centrifuge at 14 000 RPM at 4C for 10 min
#Remove supernatant to clean tube. If lysing fat, try to avoid the floating fat cake. If necessary respin to clarify
#Do a bradford assay using 1 uL of clarified lysate and label tube with correct concentration.
#Make 100 uL of SDS sample using 2X loading buffer
#Snap freeze remaining clarified lysate and store at -80
*RIPA Buffer (see [[Buffer/RIPA|RIPA]])
*Mouse Tissues
==Protocol==
#Weigh frozen tissue samples, only need 100-300 mg of tissue. If there is too much cut it off and return the extra tissue to the -80
#For fibrous tissues chop into small pieces with scissors
#Add 3 volumes of RIPA to tissue and homogenize 20x with 1 mL glass homogenizer (normally on SHC bench)
#Transfer lysate to fresh tube and keep on ice
#Clean homogenizer and lyse remaining tissues
#Centrifuge at 14 000 RPM at 4C for 10 min
#Remove supernatant to clean tube. If lysing fat, try to avoid the floating fat cake. If necessary respin to clarify
#Do a bradford assay using 1 uL of clarified lysate and label tube with correct concentration.
#Make 100 uL of SDS sample using 2X loading buffer
#Snap freeze remaining clarified lysate and store at -80