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2 protocols using anti scd1 ab39969

1

AMPK and Lipid Metabolism Signaling Pathway Analysis

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Anti-AMP-activated protein kinase (AMPK; #5831; 1:1000), anti-phospho-AMPK [Thr172] (#2535; 1:1000), anti-ACC (#3676; 1:1000), anti-phospho-ACC [Ser79] (#11818; 1:1000), anti-S6K (#2708; 1:1000), and anti-phopho-S6K [Thr389] (#9234; 1:1000) were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-SCD1 (ab39969; 1:500), and anti-actin (ab6276; 1:5000) were purchased from Abcam (Cambridge, MA, USA). Anti-microtubule-associated protein 1A/1B-light chain 3 (LC3; PM036; 1:2000) was purchased from MBL (Nagoya, Japan). Secondary anti-rabbit (#7074; 1:2000) and anti-mouse (715-035-151; 1:10000) antibodies were purchased from Cell Signaling Technology and Jackson ImmunoResearch (West Grove, PA, USA), respectively.
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2

Tanshinone IIA Regulates Lipid Metabolism

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Western blot analysis was carried out as described previously [41 (link)]. Cells were treated with vehicle or tanshinone IIA (5 and 10 μM) for 24 h. For preparation of total cellular proteins, the cells were harvested with RIPA buffer (Thermo Fisher Scientific). For nuclear-extract preparation, the cells were harvested using NE-PER nuclear and cytoplasmic extraction reagent (Thermo Fisher Scientific). The samples were separated by 10% SDS-PAGE and transferred to PVDF membranes (PerkinElmer, Boston, MA, USA). The blots were incubated with the following primary antibodies at 4 °C for 24 h: anti-FASN (A6273) (1:2000) (ABclonal, Woburn, MA, USA), anti-ACC1 (#3676) (1:1000) and anti-phospho-ACC1 (Ser79) (#3661) (1:1000) (Cell Signaling Technology, Danvers, MA), anti-SCD1 (ab39969) (1:1000) and anti-LXRα (ab41902) (1:1000) (Abcam), anti-HDAC2 (GTX112957) (1:3000) (GeneTex, Irvine, CA, USA), anti-SREBP1 (sc-13551) (1:200) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-actin (MAB1501) (1:30000) (Thermo Fisher Scientific). The blots were incubated with the appropriate HRP-conjugated secondary antibodies, and the amount of each protein was measured by Amersham ECLTM prime Western blotting detection reagent. The chemiluminescent signal was visualized using Amersham HyperfilmTM ECL film (GE Healthcare, Buckinghamshire, UK).
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