In brief, the quantities of protein in samples extracted from cardiomyocytes or the peri‐infarct region of mice were determined using a bicinchoninic acid kit. Equivalent amounts of protein were loaded in equal volumes and fractionated by SDS‐PAGE (10–15% polyacrylamide gels). GAPDH was used as the internal control. The antibodies were as follows: anti‐TGFβR3, anti‐phospho‐p38, anti‐phospho‐ERK1/2, anti‐phospho‐JNK1/2, anti‐p38, anti‐ERK1/2, anti‐JNK1/2 (1:1000 dilution; Cell Signaling Technology), and anti‐GAPDH (1:500 dilutions; Research Diagnostics).
Anti jnk1 2
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-JNK1/2 is a primary antibody that specifically recognizes the c-Jun N-terminal kinase (JNK1/2) proteins. JNK1/2 are serine/threonine protein kinases that play a key role in the regulation of various cellular processes, including cell proliferation, differentiation, and apoptosis.
Automatically generated - may contain errors
Lab products found in correlation
Anti erk1 2, by Cell Signaling Technology (14 mentions)
Anti p38, by Cell Signaling Technology (9 mentions)
Anti phospho jnk1 2, by Cell Signaling Technology (9 mentions)
Anti p jnk1 2, by Cell Signaling Technology (9 mentions)
Anti phospho erk1 2, by Cell Signaling Technology (7 mentions)
Anti akt, by Cell Signaling Technology (5 mentions)
Anti p erk1 2, by Cell Signaling Technology (5 mentions)
Anti β actin, by Merck Group (4 mentions)
Anti phospho c jun, by Cell Signaling Technology (4 mentions)
Anti β actin, by Cell Signaling Technology (4 mentions)
Anti phospho p38, by Cell Signaling Technology (3 mentions)
Anti phospho egfr, by Thermo Fisher Scientific (3 mentions)
Anti paxillin and anti c jun, by Santa Cruz Biotechnology (3 mentions)
Anti rabbit igg horseradish peroxidase linked f ab 2 1 fragment from donkey, by GE Healthcare (3 mentions)
Anti p38 mapk, by Cell Signaling Technology (3 mentions)
Anti p p38, by Cell Signaling Technology (3 mentions)
Anti phospho akt, by Cell Signaling Technology (2 mentions)
Anti egfr, by Santa Cruz Biotechnology (2 mentions)
Alexa fluor 488 conjugated anti mouse from donkey, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
26 protocols using anti jnk1 2
1
Protein Expression Analysis in Cardiac Tissue
2
Antibodies for Protein Signaling Analysis
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The following commercial antibodies were used: anti-ERK1/2, anti-phospho-ERK1/2, anti-JNK1/2, anti-phospho-JNK1/2, and anti-phospho-c-Jun (all Cell Signaling Technology, Danvers, MA, USA); anti-c-Jun, anti-phospho-Histone3, anti-p21 and anti-EGFR (Santa Cruz Biotechnology, Heidelberg, Germany); anti-phospho-EGFR and blocking anti-EGFR (Thermo Fisher Scientific, Rockford, IL, USA); anti-Paxillin (Abcam, Cambridge, MA, USA); and anti-β-Actin (Sigma-Aldrich, St Louis, MO, USA). Secondary antibodies were anti-rabbit IgG Horseradish peroxidase linked F(ab’)2 I fragment (from donkey) (GE Healthcare, GE, Little Chalfont, United Kingdom), Horseradish peroxidase linked Rat anti-mouse IgG1 (BD Pharmingen, Beckton Dickinson, Franklin Lakes, NJ, USA) and Alexa Fluor 488 conjugated anti-mouse IgG (from donkey) (Thermo Fisher Scientific, Rockford, IL, USA).
3
Antioxidant Effects of Black Rice
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Blakc rice (Oryza sativa cv. Heugjinjubyeo) was purchased from local market (Cheongju, Korea). Extracellular signal-regulated protein kinase 1/2 (ERK1/2) inhibitor (PD98059), Akt inhibitor (LY294002), Caspase-3 assay kit, 2',7'-dichlorofluorescin diacetate (DCFH-DA), tert-butyl hydroperoxide (TBHP), and 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-terazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-Akt, antiphospho-Akt, anti-ERK1/2, antiphospho-ERK1/2, anti-JNK1/2, and antiphospho-JNK1/2 were obtained from Cell Signaling Technology (Danvers, MA, USA). Fetal bovine serum (FBS), Dulbecco's modified Eagle's medium (DMEM), trypsin-EDTA, and penicillin-streptomycin were obtained from Gibco (Gaithersburg, MD, USA).
4
Antibody Staining for Signaling Proteins
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The following commercial primary antibodies were used: anti-phospho-ERK1/2, anti ERK1/2, anti-JNK1/2, anti-phospho-JNK1/2 and anti-phospho-c-Jun (all from Cell Signaling Technology, Danvers, MA, USA); anti-phospho-EGFR (Thermo Fisher Scientific, Rockford, IL, USA); anti-EGFR, anti-paxillin and anti-c-Jun (Santa Cruz Biotechnology, Heidelberg, Germany); and anti-β-actin (Sigma-Aldrich, St Louis, MO, USA). Secondary antibodies were anti-rabbit IgG Horseradish peroxidase-linked F(ab’)2 I fragment (from donkey) (GE Healthcare, GE, Little Chalfont, United Kingdom), anti-mouse IgG1 (BD Pharmingen, Beckton Dickinson, Franklin Lakes, NJ, USA), and Alexa Fluor 488 conjugated anti-mouse (from donkey) (Thermo Fisher Scientific, Rockford, IL, USA).
5
T-cell Signaling Pathway Analysis
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Jurkat T cells were stimulated with T. gondii or LPS with or without pretreatment with specific inhibitors. At the indicated time points, cells were washed with phosphate-buffered saline (PBS), and proteins were extracted using PRO-PREPTM Protein Extraction Solution (iNtRON Biotechnology, Seoul, Korea) for 15 min on ice. After centrifugation at 4°C for 15 min at 14,000 g, the supernatants were collected, and equal amounts of protein from each sample were resolved by standard SDS-PAGE and transferred to polyvinylidene difluoride membranes. The membranes were blocked with 5% skim milk in Tris-buffered saline (20 mM Tris, 137 mM NaCl, pH 7.6) containing 0.1% Tween 20 (TBST). After washing once in TBST, the membranes were incubated overnight at 4°C with anti-p-AKT, anti-AKT, anti-p-ERK1/2, anti-ERK1/2, anti-p38 MAPK, anti-p-p38 MAPK, anti-p-JNK1/2, anti-JNK1/2, and α-tubulin antibodies (all from Cell Signaling Technology) diluted in 5% bovine serum albumin in TBST. Following 3 washes in TBST, the membranes were incubated for 120 min with HRP-conjugated anti-mouse or anti-rabbit IgG (Sigma-Aldrich) diluted at 1:10,000 in incubation buffer. After extensive washing, the bound antibodies were visualized by enhanced chemiluminescence (Amersham Biosciences, Freiburg, Germany).
6
Molecular Profiling of Stem Cell Signaling
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Anti-CD44, anti-phospho-AKT, anti-AKT, anti-phospho-ERK, anti-ERK1/2, anti-phospho-p38, anti-p38, anti-phospho-SAPK/JNK, and anti-JNK1/2 antibodies were purchased from Cell Signaling Technology (Cambridge, MA, USA). Anti-phospho-JNK, anti-Oct3/4 and anti-β-actin were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-SOX2, anti-Notch2, and anti-Snail were purchased from Abcam (Cambridge, MA, USA). Anti-β-catenin was procured from BD Bioscience (Franklin Lakes, NJ, USA). For transfection, non-targeting siRNA and commercial siRNA for SOX2 or Snail or AKT were purchased from Genolution (Genolution Pharmaceuticals, Seoul, Korea). The cells were transfected with each siRNA (50 nM) for 48 h using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA), as described in the manufacturer’s procedure.
7
Molecular Signaling Pathway Immunodetection
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Anti-pSMAD2/3, anti-SMAD2/3, anti-pJNK1/2, anti-JNK1/2, anti-p-c-Jun, anti-c-Jun and anti-pCAMKII primary antibodies were purchased from Cell Signalling Technology, USA. Anti-p-ATF2 and anti-ATF2 antibodies were purchased from Santa Cruz Biotechnology, USA. Anti-CAMKII antibody was purchased from Abcam and beta-actin antibody was from Sigma-Aldrich. Anti- rabbit/mouse imunnoglobulin conjugated horseradish peroxidase antibodies were from Jackson ImmunoResearch USA. Anti-mouse/rabbit immunoglobulin conjugated Alexa Fluor 488 and Alexa Fluor 647 were from Molecular Probes, ThermoFischer Scientific. TAK1 dominant negative construct was a kind gift from Prof. K.N Balaji (MCBL, Indian Institute of Science) which was obtained from Dr. Jun Nonomiya-Tsuji (North Carolina State University, Raleigh, NC).
8
Immunoblotting Analysis of Signaling Proteins
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Equivalent amounts of protein from the samples were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and blotted onto membranes of nitrocellulose. The membranes were then incubated with anti-p65, anti-JNK1/2, anti-p38α or anti-p38β (cat no. 3034, 9252, 9218, 2339, respectively; Cell Signaling Technology, Danvers, MA, USA) or β-actin (cat no. A-1978; Sigma-Aldrich, St. Louis, MO, USA). The antibodies were afterwards detected with anti-rabbit immunoglobulin G (IgG)-HRP (cat no. 7074; Cell Signaling Technology) or with anti-mouse IgG-HRP (cat no. p0447; Dako, Glostrup, Denmark) in a standard ECL reaction (Amersham Biosciences, Piscataway, NJ, USA) according to the manufacturer's instructions.
9
Mitochondrial Dysfunction and Autophagy Assay
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Bovine serum albumin (BSA; A1595), palmitic acid (PA; P0500), Nile red (19123), p-nitrophenyl acetate (pNPA; N8130), 5′,5-dithiobis (2-nitrobenzoic acid) (DTNB; D8130), MTT cell growth assay kit (CT02), phorbol 12-myristate 13-acetate (PMA; P8139), and Rapamycin (R0395) were purchased from Sigma Aldrich (St. Louis, MO). Furthermore, 4% paraformaldehyde (BP031a) was purchased from Biosolution (Seoul, Republic of Korea). MitoSOX red mitochondrial superoxide indicator (M36008), Hoechst 33342 (H1399), and protease inhibitor cocktail (PIC; 78441) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). For immunoblotting, the following antibodies were used: anti-PON2 (Abcam, ab183718), anti-ACTB (Sigma, A5441), anti-phospho-c-Jun N-terminal kinase (JNK) 1/2 (Cell Signaling Technology, 9251), anti-JNK1/2 (Cell Signaling technology, 9252), anti-PARKIN (Cell Signaling Technology, 2132), anti-PINK1 (Novus, BC100-494), anti-BNIP3L (Cell Signaling Technology, 12396), anti-LC3B (Cell Signaling Technology, 2775), anti-SQSTM1 (Cell Signaling Technology, 5114), anti-mouse conjugated with horseradish peroxidase (Cell Signaling Technology, 7076), and anti-rabbit conjugated with horseradish peroxidase (Cell Signaling Technology, 7074).
10
Inflammatory Signaling Pathway Profiling
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TNFα (50435; Biomol Germany); IL-1β (211-11B; PeproTech); antibodies: anti-IκBα (sc-371, Santa Cruz, USA), anti-p-IκBα (9246; Cell Signaling), anti-NEMO (FL-419, Santa Cruz), anti-β Actin (sc-1616, Santa Cruz), anti-IKKβ (05–535; Millipore), anti-IKKα (14A231; Imgenex), anti-JNK1/2 (9252; Cell Signaling), anti-p-JNK1/2 (9251; Cell Signaling), anti-Rip1 (3493; Cell Signaling) anti-FLAG M2 (F3165, Sigma); anti-mCD3 (553057, BD Biosciences); anti-hCD28 (553294, BD Biosciences); anti-IgG (307-005-003, Dianova). Tris-d11 (CD4035P1, Cortecnet); DL-dithiothreitol-d10 (CD570P1, Cortecnet); D2O (151882, Aldrich); DMSO-d6 (156914, Aldrich); 15NH4Cl (CN80P100, Cortecnet); D-glucose 13C6 (CC860P20, Cortecnet); D-glucose 13C6-d7 (CCD860P20, Cortecnet).
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