Author Topic: Tissue Lysis Buffer  (Read 9600 times)

ABC123

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Tissue Lysis Buffer
« on: June 07, 2011, 11:55:25 AM »
Hello,

I am putting together a protocol to carry out protein isolation and western blotting on rat brain tissues.  I am having a difficult time deciding on the amount of lysis buffer to use for homogenizing my samples.  My samples are in the range of about 20mg, and I am afraid to add too much lysis buffer which would give me too dilute of samples, but I also want to have enough lysis buffer to be able to homogenize the tissue well.  Is there a general rule of thumb as to how much lysis buffer should be added per a given weight of tissue when working with brain tissue?

I appreciate any advice or help.  Also, I am planning on using a Tris-HCl lysis buffer, possibly containing NP-40 or Triton X-100.

Thank you!

mkvarshney

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Re: Tissue Lysis Buffer
« Reply #1 on: July 10, 2011, 06:47:52 AM »
Hi,
I am also working on rat brain tissue; doing WB and 2DGE. For tissue lysis I use 1 ml. lysis buffer per 200 mg of tissue and it works well and protein yield is also good added that you mix protease inhibitor cocktail during lysis of tissue.

Sjouke Hoving

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Re: Tissue Lysis Buffer
« Reply #2 on: October 20, 2011, 01:34:07 AM »
Hi,

This comes from the General Protocol on the sticky topic:

For a tissue sample, a rule of thumb is to take a buffer/tissue ratio of 8, in other words 1 g of tissue is lysed in 8 ml of lysis buffer.

sjouke