Lpl antibody

Protein was extracted from cultured cells by RIPA lysis buffer (Yeasen, Shanghai) with phenylmethylsulfonyl fluoride (PMSF) (1:100; Beyotime, China) and quantified with the BCA Protein Assay Kit (Beyotime). Protein lysates were separated on 15% sodium dodecyl sulfate-polyacrylamide gels and transferred to polyvinylidene difluoride membranes (0.22 μm; Millipore, Danvers, MA, USA). Then western blotting was carried out. In brief, the membranes were blocked with 5% milk at room temperature for 1 h, incubated with primary antibody overnight at 4 °C, and incubated with the appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies (1:3000; Yeasen, Shanghai) for 1 h at room temperature. Proteins were detected by ECL reagent (Millipore). The results were obtained using a Tanon imager (Tanon, Shanghai, China). The primary antibodies used were as follows: PPARγ antibody (1:1000; Abcam), AP2 antibody (1:1000; Abcam), LPL antibody (1:1000; Abcam), CEBPα antibody (1:1000; Abcam), DSP antibody (25318-1-AP, 1:500; Proteintech, Wuhan, China), β-catenin antibody (51067-2-AP, 1:1000; Proteintech), GAPDH antibody (10494-1-AP, 1:10,000; Proteintech), and HRP-labeled ACTB antibody (1:10,000; Proteintech).

The reverse transcription kit (TAKARA, article number: RR037A), SYBR kit (TAKARA, article number: RR420A), LPL antibody (Abcam, article number: ab21356), PPARγ antibody (Proteintech, article number: 16643-1-AP), internal reference GAPDH gene (Abcam, article number: ab70700), BIO-RAD Chemi Dox XRS + imaging system instrument, Eppendorf real-time PCR instrument (Mastercycler ep realplex4S), TECAN microplate reader, NBS ultra-low temperature refrigerator, SIGMA frozen high-speed centrifuge (3K30), Sanheng multipurpose electrophoresis instrument (DYY-12), and protein nucleic acid analyzer (DU650) were used.