Rabbit anti sapk jnk antibody

The samples of brain tissues were homogenized in RIPA lysis buffer with protease or phosphatase inhibitor (Roche, Basel, Switzerland) and then centrifuged (12000 rpm, 15 min, 4°C) to obtain supernatants. The total protein concentration of the supernatants was determined using the BCA assay kit (Pierce). Samples were loaded and separated on the SDS–PAGE gel (40 μg per well). The protein was transferred to the PVDF membrane (Millipore, MA, United States), which was incubated overnight with primary antibodies at 4°C and then probed with the corresponding secondary antibody. The primary antibodies were as follows: rabbit anti-AT₁R antibody (1:800 dilution, Abcam, MA, United States) (Czikora et al., 2015 (link); Wang et al., 2018 (link)), rabbit anti-SAPK/JNK antibody, mouse anti-phospho-SAPK/JNK antibody, mouse anti-ERK1/2 antibody, rabbit anti-phospho-ERK1/2 antibody, rabbit anti-p38MAPK antibody, rabbit anti-phospho-p38MAPK antibody and rabbit anti-β-tublin antibody (1:1000 dilution, Cell Signaling Technology, Danvers, MA, United States). The membranes were detected by an ECL-Plus detection kit (Tiangen, Beijing, China), and scanned using Image Quant LAS 4000 (GE Healthcare Life Sciences, CT, United States). The images were quantified using the ImageJ densitometry system.

The antibodies used were rabbit anti-p44/42 MAPK (Erk1/2) antibody #9102 (Cell Signaling Technology, Beverly, MA), rabbit anti-phospho-Erk1/2 antibody #9101 (Cell Signaling Technology), rabbit anti-phospho-p38 MAPK antibody #4511 (Cell Signaling Technology), rabbit anti-p38 MAPK antibody #8690 (Cell Signaling Technology), rabbit anti-phospho-SAPK/JNK antibody #4668 (Cell Signaling Technology), rabbit anti-SAPK/JNK antibody #9258 (Cell Signaling Technology), mouse anti-phospho-IκBα #9246 (Cell Signaling Technology), mouse anti-IκBα antibody #4814 (Cell Signaling Technology), rabbit anti-LTβR, N-Terminal antibody #SAB4501788 (Sigma-Aldrich), goat anti-LTβR antibody #L5412 (Sigma-Aldrich), normal goat IgG control #AB-108-C (R&D systems), anti-mouse-IgG HRP-linked antibody #7076 (Cell Signaling Technology), and anti-rabbit IgG-HRP-linked antibody #7074 (Cell Signaling Technology).
Equal protein loading was confirmed by probing the blot with mouse anti-α-tubulin (Sigma-Aldrich) antibody.