Srebp1 antibody

First, the whole protein was extracted by lysing cervical cancer cells with RIPA buffer before measuring the protein concentration via a BCA protein assay kit (A8020‐5, Roche, Basel, Switzerland). The cells were dissolved with SDS‐PAGE loading buffer and transferred into PVDF membranes. Then, the cells were incubated with diluted primary antibodies at 4°C after blocking with 5% skim milk dissolved in TBST for 1 hour. Primary antibodies used include HSDL2 antibody, E‐cadherin antibody, Vimentin antibody, Snail antibody, Slug antibody (Cell Signaling Technology, Boston, USA), MMP‐2 antibody (Affinity, Cincinnati, USA), FASN antibody, ACSL1 antibody, SREBP1 antibody (Proteintech) and GAPDH antibody (CWBIO). Next, goat antimouse IgG conjugated to HRP was applied as secondary antibody for 1 hour at room temperature. The cells were shortly incubated by ECL detection reagent, then detected by Imager.

The retrieved mouse liver tissue was fixed in 4% paraformaldehyde overnight, paraffin embedded and sectioned. The sections were deparaffinized, rehydrated and subjected to H & E staining and IHC staining as described.26 Briefly, the sections were subjected to deparaffinization, followed by antigen retrieval and immunostaining with different antibodies against MLXIPL (1:100‐1:200 dilution; Gen Tex; Cat#GTX30677), p‐mTOR (phospho Ser2448 1:100‐1:200 dilution; Gene Tex; Cat#GTX132803), mTOR (1:100‐1:200 dilution; and Gene Tex; Cat#GTX101557), LIPIN1 (1:100‐1:200 dilution; Abcam; Cat#ab181389), SREBP1 antibody (1:100‐1:200 dilution; Proteintech; Cat#14099‐1‐AP). Control rabbit IgG (1:200; cat. no. 011‐000‐003; Jackson ImmunoResearch Laboratories, Inc) was used as a negative control. examined and recorded under a microscope (magnification, ×400).