As previously described48 (link), proteins (30–50 μg) were separated on 4–20 % SDS-polyacrylamide gels and transferred onto PVDF membranes. The PVDF membranes were then blocked with 3% BSA in TBST followed by incubation with anti-Cytochrome c (Proteintech, IL) antibodies at 4 °C for 12 hours. After washing three times, PVDF membranes were incubated with HRP conjugated secondary antibodies (Cell Signaling; Danvers, MA, USA) for 1 hour at room temperature. Membrane-bound proteins were detected and analyzed by CCD digital imaging system and Image J analysis software (Version 1.49; NIH, USA), respectively.
Protocol detail
Quantitative Western Blot Analysis
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : Harvard–MIT Division of Health Sciences and Technology
Other organizations : Augusta University
Top 5 similar protocols
Protocol cited in 1 other protocol
Variable analysis
independent variables
- Protein amount (30–50 μg)
- Cytochrome c protein levels
- SDS-polyacrylamide gel concentration (4–20%)
- Transfer of proteins onto PVDF membranes
- Blocking of PVDF membranes with 3% BSA in TBST
- Incubation with anti-Cytochrome c antibodies at 4 °C for 12 hours
- Incubation with HRP conjugated secondary antibodies for 1 hour at room temperature
- Detection and analysis of membrane-bound proteins using CCD digital imaging system and Image J analysis software
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!