Western blot was performed, according to a previously described method (Kim et al., 2015 (link)). Protein extracts from hippocampal tissues were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Protein separation was performed using 10% polyacrylamide with 0.05% bis-acrylamide. Proteins were then transferred to nitrocellulose and the blots were probed with anti-PI3K mouse monoclonal antibody (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-Akt rabbit polyclonal antibody (1:1,000, Cell Signaling Technology Inc., Beverly, MA, USA), anti-GSK-3β rabbit polyclonal antibody (1:1,000, Santa Cruz Biotechnology), anti-synapsin I mouse monoclonal antibody (1:1,000, Santa Cruz Biotechnology), and anti-postsynaptic density-95 (PSD-95) mouse monoclonal antibody (1:1,000, Santa Cruz Biotechnology). Peroxidase anti-rabbit IgG (1:5,000, Vector Laboratories), and peroxidase anti-mouse IgG (1:10,000, Vector Laboratories) were used as the secondary antibodies. Immunoreactivity was detected by enhanced chemiluminescence (ECL) detection kit (Santa Cruz Biotechnology).
Protocol detail
Western Blot Analysis of Hippocampal Proteins
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.
anti-PI3K mouse monoclonal antibody
anti-Akt rabbit polyclonal antibody
anti-GSK-3β rabbit polyclonal antibody
Peroxidase anti-rabbit IgG
peroxidase anti-mouse IgG
ECL) detection kit
Acrylamide
Anti antibody
Anti igg
Antibodies
Chemiluminescence
Monoclonal antibody
Mouse
Nitrocellulose
Peroxidase
Pi3k
Polyacrylamide
Postsynaptic density
Proteins
Rabbit
Sodium dodecyl sulfate polyacrylamide gel electrophoresis
Synapsin i
Tissues extracts
Vector
Western blot
Corresponding Organization :
Other organizations : Hosan University, Inje University
Top 5 similar protocols
Protocol cited in 2 other protocols
Variable analysis
independent variables
- Protein extracts from hippocampal tissues
- Protein levels of PI3K, Akt, GSK-3β, synapsin I, and PSD-95
- Protein separation using 10% polyacrylamide with 0.05% bis-acrylamide
- Protein transfer to nitrocellulose membrane
- Primary antibodies: anti-PI3K, anti-Akt, anti-GSK-3β, anti-synapsin I, anti-PSD-95
- Secondary antibodies: peroxidase anti-rabbit IgG, peroxidase anti-mouse IgG
- Detection method: enhanced chemiluminescence (ECL)
- 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!