Whole cell extracts from cardiac tissue or cultured myocytes were prepared by using cell lysis buffer system (Santa Cruz). Nuclear extraction reagents (Pierce) and cytoplasmic extraction reagents (Pierce) were used to extract nuclear proteins and cytoplasmic proteins respectively according to the protocol provided by the manufacturer. Concentrations of protein samples were determined by BCA protein assay kit (Pierce). 20 μg of protein samples were loaded and then separated by SDS-PAGE. The protein samples were transferred electrically to PVDF or NC membranes which were then incubated with primary antibodies against p38 MAPK (Cell Signaling Tech, 1: 1,000), phospho-p38 MAPK (p-p38 MAPK, Cell Signaling Tech, 1: 1,000), JNK (Abcam, 1: 500), phospho-JNK (p-JNK, Abcam, 1: 500), ERK1/2 (Cell Signaling Tech, 1: 1,000), phospho-ERK1/2 (p-ERK1/2, Cell Signaling Tech, 1: 1,000), Nrf2 (Invitrogen, 1: 500), HO1 (Santa Cruz, 1: 1,000), Prx1 (Santa Cruz, 1: 1,000), GAPDH (Santa Cruz, 1: 2,000) and Lamin A (Santa Cruz, 1: 1,000) at 4°C for 10 hours. After washing with TBST, membranes were incubated with horseradish peroxidase conjugated secondary antibodies (1: 5,000) at room temperature for one hour. ECL kit (Pierce) was used to incubate the membranes which were then exposed with Gene Genius (Syngene); then the intensities of the immunoblots were analyzed by software ImageJ.
Phospho jnk p jnk
Manufactured by Abcam
Sourced in United Kingdom
Phospho-JNK (p-JNK) is a lab equipment product that detects the phosphorylated form of the c-Jun N-terminal kinase (JNK) protein. JNK is a member of the mitogen-activated protein kinase (MAPK) family and is involved in cellular responses to various stimuli, including stress and cytokines.
Automatically generated - may contain errors
Lab products found in correlation
Erk1 2, by Cell Signaling Technology (1 mentions)
P38 mapk, by Cell Signaling Technology (1 mentions)
Lamin a, by Santa Cruz Biotechnology (1 mentions)
Cell lysis buffer system, by Santa Cruz Biotechnology (1 mentions)
Phospho erk1 2 p erk1 2, by Cell Signaling Technology (1 mentions)
Genegenius, by Syngene (1 mentions)
Phospho p38 mapk p p38 mapk, by Cell Signaling Technology (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Thp 1, by American Type Culture Collection (1 mentions)
Sb203580, by Merck Group (1 mentions)
P elk 1, by Abcam (1 mentions)
Apoa 1, by Merck Group (1 mentions)
2 protocols using phospho jnk p jnk
1
Cardiac Protein Expression Analysis
2
Monocyte Differentiation and Lipid Metabolism
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Human monocyte line THP-1 was purchased from American Type Culture Collection (Rockville, MD, USA). Fetal bovine serum (FBS), phorbol-12-myristate acetate (PMA), apoA-I and SB203580 were provided by Sigma-Aldrich (St. Louis, MO, USA). Ox-LDL, acetylated low-density lipoprotein (ac-LDL) and HDL were obtained from Yiyuan Biotechnology (Guangzhou, China). Lentiviral vectors expressing asprosin (LV-Asprosin) or Elk-1S383A (serine 383 was replaced with alanine) were constructed by Genechem (Shanghai, China). Phosphorylation mutant Elk-1S383A was generated by using a QuikChange II site-directed mutagenesis kit (Stratagene, La Jolla, CA, USA) according to the manufacturer’s instructions. The mutation of Ser383 to alanine was confirmed by DNA sequencing. Rabbit antibodies against liver X receptor α (LXRα), proprotein convertase subtilisin/kexin type 9 (PCSK9), CD36, Elk-1, phospho-Elk-1 (p-Elk-1, Ser383), p-Elk-1 (Ser389), p-Elk-1 (Thr417), p38, phospho-p38 (p-38), JNK, phospho-JNK (p-JNK), ERK1/2, phospho-ERK1/2 (p-ERK1/2), ABCA1, ABCG1, histone H3 and β-actin were supplied by Abcam (Cambridge, UK). Rabbit antibody against asprosin (FineTest, Wuhan, China), mouse antibody against scavenger receptor class A (SR-A; R&D systems, Minneapolis, MN, USA), and horseradish peroxidase (HRP)-conjugated goat anti-rabbit or mouse IgG (H + L) (Beyotime, Shanghai, China) were obtained as indicated.
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