Ultrasonic bath for cell lysis - (Jan/17/2012 )

Could you please anybody give stepwise procedure for doing Ultrasonic bath forMammalian cell lysis??

Do I need to take only cells or cells with pbs or with lysis buffer???

After Ultrasonic bath what shd i do for BCA protein analysis???

Thank you,
Vino

You need to have the cells in some sort of buffer so that you can then have something to run on a gel (apart from cell debris). You can do quantitation using BCA by following the manufacturer's instructions (come with the kit), or by doing bradford assay...

I use an ultrasonic device called VialTweeter, which is very handy as you just place your vials with your samples (cells in buffer) into the VialTweeter and switch the device on. This type of sonicator is also more powerful as a common ultrasound bath and you can lyse up to 8 vials with samples at the same time. You may check this VialTweeter sonicator! In our lab it does a very good job

Dear Vinoth Shanmukam,

"Mammalian cell lysis".........the different methods are:-

Freeze Thaw
Probe Sonication
Ultrasonic water baths (adjustable and constant power)
Homogenisation

Each method is either more or less efficient at cell lysis.

Combined with this is the fact that every mammalian cell is either easier or harder to lyse than the other. So you have these 2 variables to think about. So you have to optimise to get the best returns after lysis.

You should always use a lysis buffer with Protease inhibitors if the resultant protein you want to work with is for westerns or for kinetic studies. The protease inhibitors will increase the chances of having intact active proteins to work with.

There are many kits available for estimating protein concentartion in your lysed cell solution.

Kindest regards

Uncle Rhombus