Cell lysates were prepared using complete lysis buffer (EMD Millipore, San Diego, CA) with protease and phosphatase inhibitor cocktails (Roche Diagnostics, Indianapolis, IN). Protein quantification was performed using DC protein assay from Bio-Rad (Hercules, and CA). Western blot analysis was performed as described previously (Goc et al., 2013 (link); Sabbineni et al., 2018 (link)). Antibodies used include stromelysin1, N-cadherin, and GAPDH from Cell Signaling Technology (Danvers, MA), αSMA, and β-actin from Sigma (St. Louis, MO), and ALK5 from Abcam (Cambridge, MA). Band densitometry was done using NIH Image J software.
Protocol detail
Western Blot Analysis of Cell Lysates
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.
complete lysis buffer
protease and phosphatase inhibitor cocktails
DC protein assay
N-cadherin
GAPDH
αSMA
β-actin
Actin
Alk5
Antibodies
Assay
Buffer
Cell
Densitometry
Diagnostics
Gapdh
N cadherin
Phosphatase
Protease
Protein
Rad protein
Western blot analysis
αsma
Corresponding Organization : Augusta University
Top 5 similar protocols
Protocol cited in 1 other protocol
Variable analysis
independent variables
- None explicitly mentioned
- Protein levels of stromelysin1, N-cadherin, GAPDH, αSMA, β-actin, and ALK5
- Lysis buffer composition (complete lysis buffer with protease and phosphatase inhibitor cocktails)
- Protein quantification method (DC protein assay from Bio-Rad)
- Western blot analysis method (as described previously in cited references)
- Positive controls: Not explicitly mentioned
- Negative controls: Not explicitly mentioned
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!