Are the saved TH sample is after centrifugation or homogenisation?

Manxi Zhao
Manxi Zhao
Sep 17, 2024
protocol Protocol: Endoplasmic Reticulum Isolation: An Optimized Approach into Cells and Mouse Liver Fractionation

According to the Fig.1, after homogenisation, centrifuge 5 min, 600 g first, then save TH sample and keep SN for next step.

So, where is the TH sample from? Before centrifugation or after cuntrifugation (supernatent) ?


Another question is How many protease inhibit Read more Read less more less

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María Isabel Hernández-Alvarez Author Answered Sep 18, 2024

University of Barcelona Barcelona

Dear Manxi Zhao,


The first step, 600 g 5 min, is only to collect the cells after the scraping. TH is collected only after the dounce homogenization, and it is directly from the mix that you obtain during this process without centrifuge.


Best,


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Raúl Ventura Author Answered Sep 25, 2024

Regarding the second question related with how much protease inhibitors are we adding, we use a Protease Inhibitor Cocktail (PIC). It should be at x1 in every buffer.

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