Mitochondrial Protein Isolation Buffer

A simple one buffer procedure for the isolation of proteins from the cytoplasm and mitochondria of cultured mammalian cell lines

  • Fast - Isolate mitochondrial proteins in less than 1 hour
  • Scalable - For variable number of cultured mammalian cells
  • Clean – Separate highly enriched mitochondrial fraction from the cytosol
  • Compatible – Isolated mitochondrial proteins can be used for Western blotting, ELISA and other assays

Fractionation of mitochondrial proteins from the cytosol by AMRESCO's Mitochondrial Protein Isolation Buffer

Figure 1. AMRESCO’s Mitochondrial Isolation Buffer separates the cytoplasmic proteins from the subcellular compartment. Mitochondrial (lanes 1, 3) and cytosolic (lanes 2, 4) proteins isolated from K562 cells using AMRESCO’s Mitochondrial Buffer. 10ug of protein was loaded in each lane and separated on a 12.5% Fluorescent Sprint Next™ Gel (M318). (A) Total protein was viewed on a transilluminator and then transferred to nitrocellulose for immunoblotting. Blots were blocked 5 minutes in 1X RapidBlock™ Solution (M325), then incubated with antibody to B-tubulin (1,2) or Endo G (C-term) (3,4) diluted in RapidBlock™. Blots were washed in TBS/0.1% Tween® 20 followed by incubation with with HRP- conjugated secondary antibody diluted in RapidBlock™. AMRESCO’s VisiGlo™ HRP Plus Chemiluminescent Substrate kit (N219-Kit) was used to detect the labeled proteins (B).

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