Immunoblot was carried out as others described.23 Briefly, the cells were lysed for 20 minutes on ice in RIPA lysis buffer containing a protease inhibitor cocktail and DMSF. The samples were subjected to 12% SDS‐PAGE and transferred to nitrocellulose membranes. Blots were probed with primary antibodies anti‐MARCH5 (Abcom, 1:1000), anti‐Lc3 (Abclone, 1:1000) anti‐KLF4 (Affinity, 1:1000), anti‐Drp1 (Affinity, 1:1000), anti‐Ubiquitin (Affinity, 1:1000) and anti‐β‐actin (TransGen, 1:2000) at 4°C overnight with gently shaking. After three times washing with PBS, the horseradish peroxidase (HRP)‐conjugated secondary antibodies were added. Antigen‐antibody complexes were visualized by enhanced chemiluminescence. Enhanced ECL TM prime detection reagent (GE) used to visualize antigen‐antibody complexes, the density quantified by Image J.
Enhanced ecl prime detection reagent
Manufactured by GE Healthcare
The Enhanced ECL™ Prime Detection Reagent is a chemiluminescent detection system used for Western blotting applications. It is designed to provide high sensitivity and a wide dynamic range for the detection of low-abundance proteins.
Automatically generated - may contain errors
2 protocols using enhanced ecl prime detection reagent
1
Immunoblot Analysis of Cellular Proteins
2
Apoptosis Pathway Protein Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed for 20 minutes on ice in RIPA lysis buffer containing a protease inhibitor cocktail and DMSF. The samples were subjected to 12% SDS-PAGE and transferred to nitrocellulose membranes. Blots were probed with primary antibodies anti-Caspase3 (Abcom, 1:1000), anti-Cyto c (Abclone, 1:1000) anti-Tubulin (A nity, 1:1000) anti-β-actin (TransGen, 1:2000) at 4°C overnight with gently shaking. After three times washing with PBS, the horseradish peroxidase (HRP)-conjugated secondary antibodies were added. Antigen antibody complexes were visualized by enhanced chemiluminescence. Enhanced ECL TM prime detection reagent (GE) used to visualize antigen-antibody complexes, the density quanti ed by Image J.
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!