For immunoblotting, cells were collected by trypsinization, washed with PBS and lysed in homogenization buffer (0.3 M sucrose, 10 mM Tris (pH 8.0), 400 mM sodium chloride, 3 mM magnesium chloride, 0.5% NP40/IGEPAL, 100 μg/mL Aprotinin+Protease inhibitor cocktail (Roche # 11 836 153 001). Proteins were separated by SDS-PAGE, transferred to a membrane (LI-COR), probed with primary antibodies overnight at 4°C, and probed with secondary antibodies (LI-COR) for 1–2 hours at room temperature. Proteins were visualized using the LI-COR Odyssey Infrared Imaging detection system. The following primary antibodies were used Id1, Id2, Id3, Id4 (195–14, 9-2-8, 17–3, 82–12, respectively, all from Biocheck), Cyclin D1 (2978, Cell Signaling), Actin (A2066, Sigma), Tubulin (T4026, Sigma). Western blot quantification was carried out using channel 700 and channel 800 intensity data from Odyssey application software version 3.0.30 (LI-COR), subtrActing blank values and normalizing to Tubulin.
Odyssey infrared imaging detection system
Manufactured by LI COR
Sourced in United States
The Odyssey Infrared Imaging detection system is a laboratory instrument designed for high-sensitivity, quantitative detection of proteins, nucleic acids, and small molecules in a variety of applications. The system utilizes near-infrared fluorescence technology to provide accurate and reliable measurement of analytes in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Aprotinin protease inhibitor cocktail, by Roche (3 mentions)
Odyssey application software version 3, by LI COR (2 mentions)
Cyclin d1, by Cell Signaling Technology (2 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Anti actin, by Merck Group (1 mentions)
Anti bcl 2, by Santa Cruz Biotechnology (1 mentions)
Anti pkg1, by Cell Signaling Technology (1 mentions)
Anti gapdh, by Abcam (1 mentions)
Anti nephrin, by Progen Biotechnik (1 mentions)
4 protocols using odyssey infrared imaging detection system
1
Immunoblotting Protocol for Protein Analysis
2
Immunoblotting Protocol for Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunoblotting, cells were collected by trypsinization, washed with PBS and lysed in homogenization buffer (0.3 M sucrose, 10 mM Tris (pH 8.0), 400 mM sodium chloride, 3 mM magnesium chloride, 0.5% NP40/IGEPAL, 100 μg/mL Aprotinin+Protease inhibitor cocktail (Roche # 11 836 153 001). Proteins were separated by SDS-PAGE, transferred to a membrane (LI-COR), probed with primary antibodies overnight at 4°C, and probed with secondary antibodies (LI-COR) for 1–2 hours at room temperature. Proteins were visualized using the LI-COR Odyssey Infrared Imaging detection system. The following primary antibodies were used Id1, Id2, Id3, Id4 (195–14, 9-2-8, 17–3, 82–12, respectively, all from Biocheck), Cyclin D1 (2978, Cell Signaling), Actin (A2066, Sigma), Tubulin (T4026, Sigma). Western blot quantification was carried out using channel 700 and channel 800 intensity data from Odyssey application software version 3.0.30 (LI-COR), subtrActing blank values and normalizing to Tubulin.
3
Western Blot Analysis of Renal Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Renal homogenates were separated by SDS-PAGE electrophoresis, blotted on nitrocellulose membrane (Bio-Rad, Hercules, CA, USA), and incubated with primary antibodies, followed by 1 h incubation with fluorescent secondary antibodies (Alexa Fluor, 680 nm). The Western blotting was analyzed and quantified by Odyssey Infrared Imaging Detection System (LICOR Biosciences, Lincoln, NE, USA). The optical densities were expressed as arbitrary units. Antibodies: anti-nephrin (Progen, Heidelberg, Germany); anti-actin (Sigma-Aldrich); anti-α1 Na-K ATPase (Hybridoma Bank, clone a6F, Iowa City, IA, USA); anti-PKG1 (Cell Signaling Technology, Danvers, MA, USA); anti-Bcl2 (Santa Cruz Biotechnology, CA, USA); anti-GAPDH (Abcam, Cambridge, UK).
4
Immunoblotting for ID Proteins Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunoblotting, cells were collected by trypsinization, washed with PBS and lysed in homogenization buffer (0.3 M sucrose, 10 mM Tris (pH 8.0), 400 mM sodium chloride, 3 mM magnesium chloride, 0.5% NP40/IGEPAL, 100 μg/mL Aprotinin+Protease inhibitor cocktail (Roche # 11 836 153 001). Proteins were separated by SDS-PAGE, transferred to a membrane (LI-COR), probed with primary antibodies overnight at 4oC, and probed with secondary antibodies (LI-COR) for 1-2 hours at room temperature. Proteins were visualized using the LI-COR Odyssey Infrared Imaging detection system. The following primary antibodies were used Id1, Id2, Id3, , respectively, all from (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprint this version posted January 6, 2020.
The copyright holder for this preprint this version posted January 6, 2020.
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!