Cells were lysed in TritonTM ×100-containing lysis buffer supplemented with both protease and phosphatase inhibitors. Approximately 50 µg of cellular proteins were recovered in the 10,000×g pellet from 106 cells. Cell lysates and neutralized, acid-eluted immunoprecipitates were separated by SDS-PAGE under reducing conditions and transferred to HybondTM ECLTM membranes (GE Healthcare). Membranes were incubated sequentially with the different primary antibodies and developed with the appropriate HRP-conjugated secondary antibody using enhanced chemiluminescence (ECL). When required, the blots were stripped following standard protocols prior to reprobing them with primary and HRP-conjugated secondary antibodies. Autoradiography films were scanned and the bands were quantified using Quantity One 4.4.0 (Bio-Rad) and ImageJ (Rasband, W.S., ImageJ, National Institutes of Health, Bethesda, Maryland, USA, http://rsb.info.nih.gov/ij/ , 1997–2004) softwares.
Hybond ecltm membranes
Manufactured by GE Healthcare
HybondTM ECLTM membranes are a type of lab equipment used for Western blotting analysis. They serve as a solid support for the transfer and immobilization of proteins separated by gel electrophoresis. The membranes are designed to provide efficient protein binding and transfer, enabling the detection and analysis of target proteins.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using hybond ecltm membranes
1
Western Blot Protein Analysis Protocol
2
Western Blot Analysis of Cellular Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed in Triton TM x100-containing lysis buffer supplemented with both protease and phosphatase inhibitors. Approximately 50 µg of cellular proteins were recovered in the 10,000xg pellet from 10 6 cells. Cell lysates and neutralized, acideluted immunoprecipitates were separated by SDS-PAGE under reducing conditions and transferred to Hybond TM ECL TM membranes (GE Healthcare). Membranes were incubated sequentially with the different primary antibodies and developed with the appropriate HRP-conjugated secondary antibody using enhanced chemiluminescence (ECL). When required, the blots were stripped following standard protocols prior to reprobing them with primary and HRP-conjugated secondary antibodies.
Autoradiography films were scanned and the bands were quantified using Quantity One 4.4.0 (Bio-Rad) and ImageJ (Rasband, W.S., ImageJ, National Institutes of Health, Bethesda, Maryland, USA, http://rsb.info.nih.gov/ij/, 1997-2004) softwares.
Autoradiography films were scanned and the bands were quantified using Quantity One 4.4.0 (Bio-Rad) and ImageJ (Rasband, W.S., ImageJ, National Institutes of Health, Bethesda, Maryland, USA, http://rsb.info.nih.gov/ij/, 1997-2004) softwares.
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!