Cell lysates were pre‐cleared. Some cell lysates were collected for western blot, and some lysates were incubated overnight with 2 μg capture antibodies (Anti‐SIRT1: ab189494, 1/30; Abcam, Cambridge, UK; Anti‐Flag: AF0036, 1/40; Beyotime Biotechnology, Shanghai, China) and 20 μL 50% protein A/G‐agarose beads (Pierce, Rockford, IL, USA) with mild rotation at 4°C. The A/G‐agarose beads were washed three times with modified radioimmunoprecipitation assay buffer, and boiled in the 2 × Laemmli buffer before sodium dodecyl‐sulfate polyacrylamide gel electrophoresis. Then, the immunoprecipitates were detected with specific antibodies against YAP (1:20, ab52771; Abcam) or acetyllysine (1:100, #9441; Cell Signaling, Danvers, MA, USA). Immunoglobulin G was used as the negative control of the precipitate.
Anti sirt1 ab189494
Manufactured by Abcam
Sourced in United Kingdom, United States
Anti‐SIRT1: ab189494 is a rabbit polyclonal antibody designed to detect SIRT1 protein. SIRT1 is a NAD+-dependent deacetylase that regulates various cellular processes, including metabolism, stress response, and aging. The antibody is suitable for use in western blotting, immunoprecipitation, and other immunoassay applications.
Automatically generated - may contain errors
2 protocols using anti sirt1 ab189494
1
Immunoprecipitation of SIRT1 and YAP
2
Molecular Mechanisms of Autophagy Regulation
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Fetal bovine serum (FBS) was purchased from Gibco (HyClone, Logan, UT, USA). Lipofectamine 2000 was acquired from Invitrogen (Waltham, MA, USA). Anti-Beclin1 (11306-1-AP), anti-LC3B (14600-1-AP), anti-PINK1 (23274-1-AP), anti-Tom20 (11802-1-AP), anti-PGC-1α (66369-1-Ig), and anti-Podocin (20384-1-AP) were purchased from Proteintech (Rosemont, IL, USA). Anti-P62 (ab109012), anti-Parkin (ab77924), anti-Desmin (ab32362), and anti-SIRT1 (ab189494) were purchased from Abcam (Boston, MA, USA). TFAM (AF0531) was obtained from Affinity Biosciences (Cincinnati, OH, USA). Anti-beta actin (anti-β-actin) and horseradish peroxidase-conjugated secondary antibodies were acquired from Beijing Zhong Shan Golden Bridge Biological Technology Co., Ltd. (Beijing, China).
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