For total protein extracts, tissues were homogenized in protein lysis buffer and centrifuged at 15,000g for 15 minutes at 4ºC. Nuclear extracts were isolated as previously described.57 (link) Proteins were separated by 8% or 10% wt/vol sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE), transferred to polyvinylidenedifluoride (PVDF) membranes and probed overnight at 4°C. The primary antibodies used were: Anti-Sirt1 (07-131, Millipore, Billerica, MA), Anti-AMPK (2532, Cell Signaling, Danvers, MA), Anti-phospho-AMPK-Thr172 (2531, Cell Signaling), Anti-AKT (9272, Cell Signaling), Anti-phospho-AKT-Ser473 (9271, Cell Signaling), Anti-Acetyl-p53-Lys379 (2570, Cell Signaling), Anti-Myosin Skeletal Slow (M8421, Sigma-Aldrich, St. Louis, MO), Anti-p53 (2524, Cell Signaling) and Anti-α-tubulin (ab4074, Abcam, Cambridge, UK). Detection was performed using the corresponding horseradish peroxidase-labeled secondary antibodies and western blotting detection reagent (ECL Plus; Amersham, Freiburg, Germany).
Anti myosin skeletal slow
Manufactured by Merck Group
Sourced in United States
Anti-Myosin Skeletal Slow is a lab equipment product used for the detection and quantification of slow-twitch skeletal muscle myosin in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti phospho ampk thr172, by Cell Signaling Technology (1 mentions)
Anti phospho akt ser473, by Cell Signaling Technology (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Anti p53, by Cell Signaling Technology (1 mentions)
Anti sirt1, by Merck Group (1 mentions)
Anti acetyl p53 lys379, by Cell Signaling Technology (1 mentions)
Anti ampk, by Cell Signaling Technology (1 mentions)
Ecl plu, by Cytiva (1 mentions)
Anti α tubulin, by Abcam (1 mentions)
Fusioncapt advance fx7, by Vilber (1 mentions)
2 protocols using anti myosin skeletal slow
1
Western Blot Analysis of Protein Extracts
2
Quantifying Skeletal Muscle Protein Isoforms
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Cells were lysed with buffer containing 50 mM Tris (pH 8.0), 150 mM NaCl, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS, 5 mM EDTA, and protease inhibitor cocktail (cOmpleteTM, 11873580001, Roche, Basel, Switzerland). Extracts were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to 0.45 µm pore size nitrocellulose membrane (1620115, Bio-Rad, Hercules, CA, USA). Western blot was performed using standard procedures, with blocking in 5% skimmed milk for 1 h and washes in PBS containing 0.05% Tween 20. Membranes were probed at 4 °C overnight with target antibodies: Anti-skeletal myosin (FAST) (M4276, Sigma-Aldrich, St. Louis, MO, USA), anti-myosin (Skeletal, Slow) (M8421, Sigma-Aldrich, St. Louis, MO, USA), and myosin heavy chain (MF20, R&D Systems, Minneapolis, MN, USA). The blots were revealed by secondary HPR-conjugated antibodies (#1706516, Bio-Rad, Hercules, CA, USA), and chemiluminescense was detected using Fusion Fix imaging system (Vilber Lourmat, Marne La Vallee, France), and analyzed with FusionCapt Advance FX7.
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