Whole fresh retinas were rapidly isolated and homogenized in ice-cold lysis buffer: 150 mM NaCl, 20 mM Tris, pH 8.0, 1% NP-40, 0.5% Na deoxycholate, 0.1% SDS, 1 mM EDTA, supplemented with 2 mM NaVO3, and protease and phosphatase inhibitors. Protein samples were resolved on SDS polyacrylamide gels and transferred to nitrocellulose membranes (Bio-Rad Life Science, Mississauga, ON). Blots were incubated with each of the following antibodies: pAMPKThr172 (0.027 μg/ml, Sigma), total AMPK (1:1000, Sigma), phospho-LKB1 (Ser248, 0.183 μg/ml, Sigma), total LKB1 (0.024 μg/ml, Sigma), or β-actin (0.5 μg/ml, Sigma-Aldrich), followed by anti-rabbit or anti-mouse peroxidase-linked secondary antibodies (0.5 μg/ml, GE Healthcare, Mississauga, ON). Blots were developed with a chemiluminescence reagent (ECL, Amersham Biosciences) followed by exposure of blots to ChemiDoc MP System (Bio-Rad Life Science). Analysis was performed using densitometric software (Bio-Rad Life Science) and three independent western blots were carried out using retinal samples from distinct experimental groups.
Anti rabbit or anti mouse peroxidase linked secondary antibodies
Manufactured by GE Healthcare
Anti-rabbit or anti-mouse peroxidase-linked secondary antibodies are laboratory reagents used in immunoassays and immunodetection techniques. They are designed to bind and detect primary antibodies raised against rabbit or mouse antigens, respectively. The peroxidase enzyme label allows for colorimetric or chemiluminescent detection of target proteins.
Automatically generated - may contain errors
Lab products found in correlation
β actin, by Merck Group (2 mentions)
Nitrocellulose membrane, by Bio-Rad (2 mentions)
Enhanced chemi luminescence (ecl), by Cytiva (1 mentions)
Chemidoc mp system, by Bio-Rad (1 mentions)
Densitometric software, by Bio-Rad (1 mentions)
Rictor, by Thermo Fisher Scientific (1 mentions)
Raptor, by Abcam (1 mentions)
Total akt, by Cell Signaling Technology (1 mentions)
Phospho akt ser473, by Cell Signaling Technology (1 mentions)
2 protocols using anti rabbit or anti mouse peroxidase linked secondary antibodies
1
Retinal AMPK Signaling Pathway Analysis
2
Retinal Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole retinas were rapidly isolated and homogenized in ice-cold lysis buffer: 150 mM NaCl, 20 mM Tris, pH 8.0, 1% NP-40, 0.5% Na deoxycholate, 0.1% SDS, 1 mM EDTA, supplemented with 2 mM NaVO3, and protease and phosphatase inhibitors. Protein samples were resolved on SDS polyacrylamide gels and transferred to nitrocellulose membranes (Bio-Rad Life Science). Blots were incubated with each of the following antibodies: phospho-Akt (Ser473, 1:2000, Cell Signaling Technology), total Akt (1:2000, Cell Signaling Technology), Rictor (1:2000, Thermo Fisher Scientific), Raptor (0.96 μg/ml, Abcam), or β-actin (0.5 μg/ml, Sigma-Aldrich), followed by anti-rabbit or anti-mouse peroxidase-linked secondary antibodies (0.5 μg/ml, GE Healthcare). Densitometric analysis was performed using ImageJ (http://imagej.nih.gov/ij/ ) on scanned autoradiographic films obtained from five independent western blots, each carried out using retinal samples from different experimental groups.
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!