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Just a quick question to see if you do any cell fractionation kits for samples from whole tissue?
I've noticed ab109718 and ab109719 but they only mention cultured cells?
Asked on Aug 17 2012
Thank you fro your enquiry.
I am sorry to confirm we would not recommend to use ab109718 Cell Fractionation Kit - HTor ab109719 Cell Fractionation Kit Standardfor isolation of mitochondria from tissue. They are designed for cell culture single cell suspension samples only. The issue with solid tissue is in the difficulty to prepare the starting material for the fractionation: single cell suspension without breaking the cells.
If you want to isolate mitochondria, then I can recommend to consider ab110168 or ab110169 instead. These are designed for mitochondrial isolation from tissue.
https://www.abcam.com/index.html?datasheet=110168 (or use the following: https://www.abcam.com/index.html?datasheet=110168).
https://www.abcam.com/index.html?datasheet=110169 (or use the following: https://www.abcam.com/index.html?datasheet=110169).
The important steps to consider from the protocol provided on the datasheetare as follows:
Spin 1 isolates the unbroken cells and nucleiSpin 2 isolates the mitochondria. The remaining supernatant after spin 2 contains mostly cytosolic proteins –the protocol suggests to save this supernatant sample (step 12) and these could be used.
There are wash steps to clean up the mitochondria (Steps 13-15) However the supernatant sample could also be washed by spinning at 12000g repeatedly and collecting the supernatant each time.
I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.
Answered on Aug 17 2012