PQ was purchased from Sigma-Aldrich (St. Louis, MO, USA). CB2R-agonist JWH133 was obtained from Enzo Life Sciences Ltd. (UK). Antibodies to p38MAPK, phosphor-p38MAPK, JNK1/2, phosphor-JNK1/2, ERK1/2, phosphor-ERK1/2, NF-κBp65, IκB-α, phosphor-IκBα, and β-actin were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA). Rabbit anti-CB2 receptor polyclonal antibody was purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA). The rat MPO determination kit was purchased from Jiancheng Bioengineering Institute of Nanjing (Nanjing, China). The ELISA kits for TNF-α and IL-1β were purchased from R&D system (R&D, USA).
Phosphor jnk1 2
Manufactured by Cell Signaling Technology
Sourced in United States
Phosphor-JNK1/2 is a lab equipment product that detects and quantifies the phosphorylation of JNK1 and JNK2 proteins. It is used to measure the activation status of these signaling molecules in cellular samples.
Automatically generated - may contain errors
Lab products found in correlation
Nf κb p65, by Cell Signaling Technology (3 mentions)
Jnk1 2, by Cell Signaling Technology (3 mentions)
Erk1 2, by Cell Signaling Technology (2 mentions)
Phosphor erk1 2, by Cell Signaling Technology (2 mentions)
Bicinchoninic acid protein assay kit, by Beyotime (2 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Phosphor iκbα, by Cell Signaling Technology (1 mentions)
Elisa kits for tnf α and il 1β, by R&D Systems (1 mentions)
Total cdk2, by Cell Signaling Technology (1 mentions)
Total cdc2, by Cell Signaling Technology (1 mentions)
Total erk1 2, by Promega (1 mentions)
Anti α tubulin, by Merck Group (1 mentions)
Phosphor cdc2, by Cell Signaling Technology (1 mentions)
Phospho p38, by Promega (1 mentions)
Phospho erk1 2, by Cell Signaling Technology (1 mentions)
Phosphor nf κb p65, by Cell Signaling Technology (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Goat anti rabbit secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Ripa buffer, by Roche (1 mentions)
Lysis buffer, by Beyotime (1 mentions)
4 protocols using phosphor jnk1 2
1
Cannabinoid Receptor-Mediated Signaling Pathways
2
Western Blot Analysis of Cell Signaling
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A Western blot analysis of whole-cell lysates was conducted as previously described [17 (link)]. The blots were probed with the following primary antibodies: phosphor-CDK2, total CDK2, CDK3, phosphor-CDC2, total CDC2, phospho-ERK1/2, phosphor-JNK1/2 (all from Cell Signaling), CDK4 (Santa Cruz Biotechnology), total p38, total ERK1/2, total JNK1/2, phospho-p38, total p38 MAPK (all from Promega), anti α-tubulin (Sigma) and anti-LC3 (MBL). Immunoreactive proteins were detected as previously described [17 (link)].
3
Investigating LPS-Induced Inflammatory Signaling
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For this stage, 1.5 × 106 of RAW264.7 cells were seeded in the 6 cm culture dish. Two days later, the cells were pretreated with Farrerol for 1 h and then stimulated with LPS for 1 h. RAW264.7 cells were lysed by RIPA buffer (Roche Diagnostics, Swissland) on ice for 15 min. A bicinchoninic acid protein assay kit (Beyotime Inst. Biotech, Shanghai, China) was used to measure protein concentration in the samples. Subsequently, 50 μg proteins were separated through 10% or 12% SDS-PAGE and then transferred to PVDF membranes. After blocking in 5% non-fat milk dissolved in Tris buffered saline for 1 h, the membranes were incubated in primary antibodies against iNOS (1:4000), COX-2 (1:2000), phosphor-ERK1/2 (1:1000), ERK1/2 (1:1000), phosphor-p38 (1:1000), p38 (1:2000), phosphor-JNK1/2 (1:2000), JNK 1/2 (1:2000), phosphor-NF-κB p65 (1:2000), NF-κB p65 (1:2000) (Cell Signaling Technology, Danvers, MA, USA), and β-actin (1:2000) (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) at 4 °C overnight. Subsequently, the membranes were washed for three times and incubated with the secondary goat anti-rabbit antibody (1:2000; Santa Cruz, CA, USA) for 1 h. By enhancing chemiluminescence, membranes could be visualized. (ECL kit; Applygen Inst. Biotech, Beijing, China).
4
Galangin Modulates Neuroinflammation Pathways
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Complete Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, Grand Island, NY, USA) was replaced with serum-free DMEM medium to starve the BV-2 cells for 4 h. Cells were pretreated with various concentrations of galangin for 1 h, followed by treatment with LPS (1 μg/mL) for different times. After the last behavioral test, the SNs of the rats were rapidly dissected out, frozen, and stored in a deep freezer at −80 °C until the assays were performed. The BV-2 cells and the rat SNs were lysed with lysis buffer for 30 min (Beyotime Inst. Biotech, Beijing, China). After centrifugation at 12,000× g at 4 °C, the supernatant protein was quantified with a bicinchoninic acid protein assay kit (Beyotime Inst. Biotech, Beijing, China). The detailed process is referenced in our previous work [24 (link)]. The source of the commercially available antibodies used for the western blot were OX-42 (1:1000), TH (1:1000), COX-2 (1:1000), iNOS (1:2000) (Abcam, Cambridge, CA, USA), phosphor-NF-κB p65 (1:1000), NF-κB p65 (1:1000), phosphor-AKT (1:2000), AKT (1:2000), phosphor-p38 (1:2000), p38 (1:1000), phosphor-ERK1/2 (1:2000), ERK1/2 (1:2000), phosphor-JNK1/2 (1:1000), JNK1/2 (1:2000), (Cell Signaling Technology, Danvers, MA, USA), and β-tubulin (1:2000) (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA).
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