Na k atpase antibody

20 μg protein were separated by 8.0% SDS-polyacrylamide gel electrophoresis and transferred to a polyvinylidene fluoride (PVDF) membrane. After transfer, the PVDF membranes were blocked with 5% (w/v) skim milk in Tris-buffered saline (TBST) [10 mM Tris HCl (pH 7.5), 150 mM NaCl, and 0.1% Tween-20] for 1 h at room temperature (RT). The PVDF membranes were then blotted with rabbit polyclonal PKCγ antibody (1 : 2000, Abcam Co., Ltd., USA) or rabbit monoclonal PKCε antibody (1 : 1000, Santa Cruz, USA), or rabbit TRPV1 polyclonal antibody (1 : 1000, Abcam Co., Ltd., USA), or rabbit p-TRPV1 polyclonal antibody (1 : 1000, Abcam Co., Ltd., USA), or rabbit monoclonal GAPDH antibody (1 : 10000, Abcam Co., Ltd., USA), or rabbit polyclonal Na⁺-K⁺-ATPase antibody (1 : 1000, Cell Signaling Technology, USA) for the DRG and SCDH samples at 4°C overnight in 5% skim milk. Then, the samples were incubated with HRP conjugated goat anti-rabbit antibody for 2 h at RT, the bands were detected by chemiluminescence using Western ECL kit (Abcam Co., Ltd., USA), and densitometry was performed by Image Quant LAS 4000 (GE, USA). The integrated optical density (IOD) of the bands was analyzed by Image Quant TL7.0 Analysis Software (GE, USA).

Proteins were extracted from liver tissues and HepG2 cells using a cell membrane protein and plasma protein extraction kit (Beyotime, P0033), and the following antibodies were used for Western blot (WB) analyses: BSEP polyclonal antibody (PA5‐78690, Invitrogen, Carlsbad, CA, USA); caveolin‐1 antibody (Cell Signaling Technology, Inc., Danvers, MA, USA); anti‐Hax‐1 antibody (Ab137613, Abcam plc., Cambridge, UK); PKCα antibody (Cell Signaling Technology, Inc.); phospho‐PKCα/β II (Thr638/641) antibody (Cell Signaling Technology, Inc.); mouse anti‐GAPDH antibody (Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China); Na,K‐ATPase antibody (Cell Signaling Technology, Inc.); and mouse anti‐β actin antibody (Zhongshan Golden Bridge Biotechnology Co., Ltd.). Protein expression was normalized to β‐actin, GAPDH and Na⁺, K⁺‐ATPase. Densitometry analyses were performed using the ImageJ software.
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