Aug 07, 2015
Whole protein extraction from tissues
- Vivian Liu
Protocol Citation: Vivian Liu 2015. Whole protein extraction from tissues. protocols.io https://dx.doi.org/10.17504/protocols.io.dkx4xm
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
Created: August 06, 2015
Last Modified: March 28, 2018
Protocol Integer ID: 1399
Abstract
This protocol is for whole protein extraction from tissues like kidney, liver ,musscle etc. Histone proteins which are tightly twisted on genomic DNA can also be extracted with this protocol.
Before start
ULTRA-TURRAX Dispenser T10 basis S25 (IKA)
Rotating mixer
Bench top centrifuge
Pre-cool lysis buffer on ice. Add protease inhibitors just before using.
Add 10 fold lysis buffer to tissues (1ml lysis buffer / 100mg tissue)
Note
Homogenize tissues with dispenser ( working time 10s / rest time 20s, repeat 5 times) until no visible tissue blocks.
Mix samples on a rotating mixer at 4 °C for 2h.
Spin @ 16,000g, 10min, 4°C. If necessary, centrifuge again.
Save supernatant. Determine protein conc. .
Make aliquots and store in -80°C.