Mouse skeletal muscle-derived C2C12 myoblasts (GNM26) were purchased from the Chinese Academy of Sciences (Shanghai, China). Fetal bovine serum (FBS) was purchased from Clark Bioscience (Richmond, VA, USA). Horse serum was purchased from GIBCO (Life Technologies, Carlsbad, CA, USA). DEX was purchased from Sigma (St. Louis, MO, USA). The lactic dehydrogenase (LDH) Biochemical Analysis Kit was purchased from Jiancheng Bioengineering Institute (Nanjing, China). Fluo-3/AM and the BCA Protein Assay Kit were purchased from Beyotime Biotechnology (Shanghai, China). Protease/phosphatase inhibitor cocktails were purchased from Roche (Basel, Switzerland). Primary antibodies against myogenin, MuRF 1, atrogin-1, phosphorylated AMPK (p-AMPK), AMPK, phosphorylated PI3K (p-PI3K), PI3K, phosphorylated Akt (p-Akt), Akt, GLUT4, and GAPDH were obtained from Cell Signaling Technology (Beverly, MA, USA). p-AS160 was obtained from OmnimABs (Alhambra, CA, USA). Chemiluminescence reagents were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
Atrogin 1
Manufactured by Cell Signaling Technology
Sourced in United States
Atrogin-1 is a protein that plays a key role in the ubiquitin-proteasome system, which is responsible for the degradation of specific proteins within cells. It functions as an E3 ubiquitin ligase, marking target proteins for destruction by the proteasome.
Automatically generated - may contain errors
Lab products found in correlation
Murf1, by Cell Signaling Technology (2 mentions)
4e bp1, by Cell Signaling Technology (2 mentions)
Fetal bovine serum (fbs), by Clark Bioscience (1 mentions)
Glut4, by Cell Signaling Technology (1 mentions)
Protease phosphatase inhibitor cocktail, by Roche (1 mentions)
Chemiluminescence reagent, by Santa Cruz Biotechnology (1 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Fluo 3 am, by Beyotime (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Myogenin, by Cell Signaling Technology (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Murf1, by Novus Biologicals (1 mentions)
Ps256 foxo1, by Cell Signaling Technology (1 mentions)
Ps473 akt, by Cell Signaling Technology (1 mentions)
Ps2448 mtor, by Cell Signaling Technology (1 mentions)
Pt172 ampk, by Cell Signaling Technology (1 mentions)
Foxo1, by Cell Signaling Technology (1 mentions)
Colorimetric protein assay kit, by Bio-Rad (1 mentions)
Chemidoc, by Bio-Rad (1 mentions)
3 protocols using atrogin 1
1
Myoblast Differentiation and Metabolic Regulation
2
Molecular Markers of Muscle Aging
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Primary antibodies were obtained as follows: LC3B, tubulin, and MuRF1 antibodies were acquired from Novus Biologicals (Centennial, CO, USA), Abcam (Cambridge, UK), and Santa Cruz Biotechnology (Dallas, TX, USA), respectively; mTOR, pS2448-mTOR, Akt, pS473-Akt, ribosomal S6, pS235/236-ribosomal S6, 4EBP1, pS65-4EBP1, FoxO1, pS256-FoxO1, Atrogin-1, ULK1, pS757-ULK1, AMPK, and pT172-AMPK were obtained from Cell Signaling Technology (Danvers, MA, USA). All secondary antibodies were procured from Jackson Immuno Research Laboratories, Inc. (West Grove, PA, USA). The intercostal, diaphragm, and gastrocnemius muscles from male Sprague Dawley rats, either 6-month-old or 20-month-old were obtained from the Aging Tissue Bank (Pusan University, Pusan, Korea) [49 (link)].
3
Muscle Protein Signaling Pathway Analysis
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The soleus fibers were homogenized using a lysis buffer and centrifuged at 13,000 rpm for 20 minutes. Protein concentration was measured via colorimetric protein assay kit (Bio-Rad, Hercules, CA, USA). Proteins (30 µg) were separated on sodium dodecyl sulfate-polyacrylamide gels and transferred onto a nitrocellulose membrane on ice. The membranes were incubated using 5% skim milk with Tris-buffered saline which is containing 0.1% Tween-20 (TBS-T) and then, incubated overnight at 4°C with the following primary antibodies: GAPDH, Bcl-2, Bax, cytochrome c (Santa Cruz Biotechnology, Santa Cruz, CA, USA), Akt, p-AKT, mTOR, p-mTOR, p70S6k, p-p70S6k, 4EBP1, p-4E-BP1, FOXO3, MuRF1, and Atrogin-1 (Cell Signaling Technology, Danvers, MS, USA). The membranes were incubated for 1 hour with secondary antibodies: horseradish peroxidase (HRP)-conjugated anti-mouse (Santa Cruz Biotechnology) or goat anti-rabbit IgG-heavy and light chain antibody HRP conjugated. After washing 3 times in TBS-T, an enhanced chemiluminescence detection kit (Thermo Fisher Scientific, Waltham, MA, USA) was used to detect the band. Protein bands was expressed using a ChemiDoc (Bio-Rad). All protein levels were calculated with GAPDH as a ratio.
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