Mitochondrial protein complexes were isolated from heart mitochondria as previously described (Wittig et al., 2006 (link)). Briefly, 400 µg mitochondria was solubilized in buffer (50 mM sodium chloride, 50 mM imizadole HCl, 50 mM Bis-Tris, 2 mM 6-aminohexanoic acid, 1 mM EDTA) followed by digestion with 20% digitonin (Sigma). Samples were loaded in a NativePAGE 4–16% gel (Invitrogen) with dark blue cathode buffer and clear anode buffer. Samples were run at 150V for 2 hours and then the gel was rinsed in water and stained with SimplyBlue SafeStain (ThermoFisher Scientific) for 1 hour and rinsed again. The gel was then incubated overnight in 700 mM NaCl (Sigma) to remove excess stain and imaged the following day using a light box.
Nativepage 4 16 gel
Manufactured by Thermo Fisher Scientific
The NativePAGE 4–16% gel is a laboratory equipment product designed for the analysis of native protein complexes. It is a pre-cast polyacrylamide gel with a gradient concentration range of 4–16% acrylamide. The gel is used to separate and analyze protein complexes under non-denaturing conditions, preserving their native structure and interactions.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using nativepage 4 16 gel
1
Isolation and Analysis of Mitochondrial Protein Complexes
2
Isolation and Analysis of Mitochondrial Protein Complexes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mitochondrial protein complexes were isolated from heart mitochondria as previously described (Wittig et al., 2006 (link)). Briefly, 400 µg mitochondria was solubilized in buffer (50 mM sodium chloride, 50 mM imizadole HCl, 50 mM Bis-Tris, 2 mM 6-aminohexanoic acid, 1 mM EDTA) followed by digestion with 20% digitonin (Sigma). Samples were loaded in a NativePAGE 4–16% gel (Invitrogen) with dark blue cathode buffer and clear anode buffer. Samples were run at 150V for 2 hours and then the gel was rinsed in water and stained with SimplyBlue SafeStain (ThermoFisher Scientific) for 1 hour and rinsed again. The gel was then incubated overnight in 700 mM NaCl (Sigma) to remove excess stain and imaged the following day using a light box.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!