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Products and Related Resources for Fibrosis SARS-CoV-2 Research
Information and case study data for the PTMScan Direct Ser/Thr Kinases Service, which allows for the targeted screening of a defined set of protein modification sites.
In-house, we tested AMPK Control Cell Extracts #9158 and mouse liver extract, blotting with AMPKα2 Antibody #2757 (approved for human and monkey) and a competitor AMPKα2 antibody (approved for mouse reactivity). We observed no signal with AMPKα2 Antibody #2757 in the mouse extracts, confirming AMPKα2 Antibody #2757 is not approved for mouse reactivity. Blotting with the competitor antibody we observed very weak signal in AMPK Control Cell Extracts #9158 and strong signal in mouse liver extract; therefore, suggesting AMPK Control Cell Extracts #9158 may not be an appropriate control for measuring AMPKα2 levels.
The Ser485 site detected by our Phospho-AMPKα1 (Ser485) (45F5) Rabbit mAb #2537 has been identified as an inhibitory phosphosite of APMKα1 and will be phosphorylated to a lesser degree when AMPK is activated in response to nutrient-deprived conditions (e.g., serum or glucose starvation). Therefore, you should see a decrease in signal intensity with #2537 in serum-starved samples, such as the positive extract included in our AMPK Control Cell Extracts #9158.
Several different assays can be used by scientists to measure metabolism in a variety of contexts. These include methods to determine metabolic rates, key signaling pathways, and environmental cues.
While we know from our experiments that our AMPKα Antibody #2532 can detect both the alpha1 and alpha2 isoforms of the catalytic subunit, these should both run above 60kDa. We do occasionally see some non-specific banding around 50 kDa or lower. However, this appears to be very sample dependent and can often be titrated out by loading less lysate. The specific AMPKα signal should be a prominent band around 62kDa.
The specificity of our Phospho-AMPKα (Thr172) (40H9) Rabbit mAb #2535 has been confirmed as follows:1) see the western image on the product webpage at https://www.cellsignal.com/products/primary-antibodies/phospho-ampka-thr172-40h9-rabbit-mab/2535. The specificity of #2535 has been confirmed on a binary model (i.e., on extracts from C2C12 cells, untreated or oligomycin-treated (0.5 µM). 2) shRNAPMID: 28553939 (https://pubmed.ncbi.nlm.nih.gov/28553939/) Figure 1b3) CRISPR/Cas9 PMID: 30415923 (https://pubmed.ncbi.nlm.nih.gov/30415923/) Figure 3A
The three main purposes of metabolism are the conversion of food to energy; the conversion of nutrients to proteins, carbohydrates, lipids, and nucleic acids; and the elimination of nitrogenous wastes. There is a strong correlation between metabolic chan
Explore the AMPK pathway and its role in cellular energy homeostasis. Uncover the mechanisms of energy regulation. Learn more here.
Webinar featuring Reuben Shaw, Professor at the Molecular and Cell Biology Laboratory, Deputy Director of Salk Cancer Center, Salk Institute for Biological Studies.
Overview of Cellular Metabolism pathways, antibodies and related reagents, interactive pathway diagrams, and other technical resources
In this webinar, Dr. Reuben Shaw will explore new findings in the field of AMPK and Autophagy signaling.
Discover the autophagy pathway and its crucial role in cellular homeostasis. Learn more and uncover the mechanisms of cellular self-degradation here.
Expert-reviewed interactive pathway providing a current overview of the Warburg Effect.
Expert-reviewed interactive pathway providing a current overview of Regulation of eIF4E and p70 S6K.
Tumor cells adopt novel mechanisms to acquire nutrients and reprogram metabolic pathways to meet their increased bioenergetics demands. CST provides tools to detect and measure key metabolic changes.
Autophagy helps normal cells maintain homeostasis and acts as a survival mechanism in response to stress, where ULK1 plays an important role in signaling.
The specificity of our AMPKα Antibody #2532 has been confirmed as follows:1) siRNAPMID: 28008135 (https://pubmed.ncbi.nlm.nih.gov/28008135/) Figure 2A2) shRNAPMID: 28553939 (https://pubmed.ncbi.nlm.nih.gov/28553939/) Figure 1b3) CRISPR/Cas9 PMID: 30415923 (https://pubmed.ncbi.nlm.nih.gov/30415923/) Figure 3A
The PI3K / Akt Substrates Table provides a comprehensive list of demonstrated downstream targets of Akt phosphorylation.
If CST recommended buffers are being used for western blotting and high background or additional bands persist when probing for phospho-AMPK (Thr172), the issue could be related to the lysate sample. In general, tissue samples can be prone to high background by western blotting when compared to cultured cell lines, and liver tissue samples (a high expressing model for AMPK) in particular tend to show high background in the form of multiple bands when probing for phospho-AMPK. Sonication and the use of proper buffers for western blotting can help to alleviate this issue in some instances. For further technical support and troubleshooting help, please reach out to a scientist at [email protected].
Cellular metabolism encompasses all of the chemical processes that occur within cells to maintain homeostasis and define their energetic status. These include anabolism and catabolism.
Overview of autophagy signaling, antibodies and related reagents, interactive pathway diagrams, and technical resources for autophagy research.
Discover the insulin signaling pathway and its role in glucose metabolism & diabetes. Learn more about insulin action here.
Cell Signaling Technology pathways by research area
Expert-reviewed interactive pathway providing a current overview of ErbB/HER Signaling.
The regulation of ER stress and autophagy during viral infection is an important factor in the balance of virus and host survival.