Liver samples were collected from six fish from each treatment. Western blotting was conducted using RIPA buffer (NCM Biotech, China) according to the method described previously (Li et al. 2020b (link)). The anti-CPT1a (1:800, 15184-1-AP, Proteintech), anti-LXRα/β (1:1000, sc-271064, Santa Cruz Biotechnology), anti-SREBP1c (1:1000, 66875-1-Ig, Proteintech), anti-LC3 (1:800, #4107, CST), anti-LAL (1:500, Custom Antibody Services from HuaAn Biotech, China), anti-IRβ (1:800, 20433-1-AP, Proteintech), anti-PI3K (1:800, CY6915, Abways), anti-p-PI3K (Tyr458/199) (1:800, #4228, CST), anti-AKT (1:800, #4091S, CST), anti-p-AKT (1:800, #4260S, CST), anti-mTORC1 (1:1000, #2972S, CST), anti-p-mTORC1 (1:1000, #2971S, CST), anti-S6 (1:1000, #2217S, CST), anti-p-S6 (1:800, AY0578, Abways), anti-GAPDH (1:3000, AB0036, Abways) and anti-α-Tubulin (1:1000, M1501-1, HuaAn Biotech) were used to measure the expressions of targeted proteins. The Western blotting images were obtained using an Odyssey CLx Imager (Licor, USA).
Sc 271064
Manufactured by Santa Cruz Biotechnology
Sourced in China
Sc-271064 is a laboratory instrument produced by Santa Cruz Biotechnology. It is designed for conducting various scientific experiments and analyses. The core function of this product is to facilitate research activities in a laboratory setting.
Automatically generated - may contain errors
Lab products found in correlation
Ab0036, by Abways (1 mentions)
Odyssey clx imager, by LI COR (1 mentions)
Ab218528, by Abcam (1 mentions)
Sc 69778, by Santa Cruz Biotechnology (1 mentions)
Supersignal chemiluminescent reagent, by Thermo Fisher Scientific (1 mentions)
Sc 2004, by Santa Cruz Biotechnology (1 mentions)
Bca assay, by Beyotime (1 mentions)
2 protocols using sc 271064
1
Western Blotting Analysis of Liver Proteins
2
Western Blot Analysis of ABCG1, IRAK-1, RXR, and LXR
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Cells were harvested and lysed in RIPA buffer. Protein content was measured using a BCA assay (Beyotime Institute of Biotechnology). Protein (50 µg) was separated via 10% SDS-PAGE and transferred to poly-vinylidene difluoride membranes, which were washed in Tris-buffered saline with Tween-20 (TBST) for three times and blocked with 5% non-fat in TBST for 1 h at room temperature. Subsequently, the membranes were incubated with antibodies against ABCG1 (ab218528, 1:1,000, Abcam), IRAK-1 (sc-515512, 1:1,000, Santa Cruz Biotechnology, Inc.), retinoid X receptor (RXR, sc-46659, 1:1,000, Santa Cruz Biotechnology, Inc.), liver X receptor (LXR, sc-271064, 1:1,000, Santa Cruz Biotechnology, Inc.) and GAPDH (sc-69778, 1:5,000, Santa Cruz Biotechnology, Inc.) at 4°C overnight. The membrane was washed three times with TBST for 15 min and incubated with a horseradish peroxidase-conjugated secondary antibody (sc-2004, 1:5,000, Santa Cruz Biotechnology, Inc.) at 37°C for 1 h. Proteins were detected by Super Signal chemiluminescent reagent (Thermo Fisher Scientific, Inc.) and measured using Flowjo 7.6.1 (FlowJo, LLC).
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