Sal was procured from Shanghai Tauto Biotechnology (Shanghai, China), the alanine transaminase (ALT), aspartate aminotransferase (AST), and hydroxyproline (Hyp) detection kits were acquired from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). FN, type I collagen (Col I), Bax and Bcl-2 antibodies were obtained from Abcam (Cambridge, United Kingdom). JNK, p-JNK, PARP, and cleaved caspase 3 antibodies were purchased from Cell Signaling Technology (Massachusetts, United States). SphK1, SphK2, S1PR1, S1PR2, S1PR3, CD9, and tumor susceptibility gene 101 (TSG101) antibodies were procured from ImmunoWay Biotechnology Company (Suzhou, China). Antibody against GAPDH was provided by Proteintech (Wuhan, China). TdT-mediated dUTP nick end labeling (TUNEL) Apoptosis Assay Kit, 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DPAI), and Cy3-labeled goat anti-rabbit IgG were all bought from Beyotime Institute of Biotechnology (Shanghai, China). The S1P Assay Kit (S1P-ELISA) was delivered by Echelon Biosciences (Salt Lake City, United States).
P jnk jnk
Manufactured by Cell Signaling Technology
Sourced in United States
The P-JNK/JNK product is a laboratory tool designed to detect and quantify the levels of both phosphorylated and total JNK (c-Jun N-terminal kinase) proteins in cellular samples. JNK is a member of the mitogen-activated protein kinase (MAPK) family and plays a crucial role in various cellular processes, including stress response, proliferation, and apoptosis. This product enables researchers to analyze the activation status of the JNK signaling pathway, which is important for understanding cellular signaling mechanisms.
Automatically generated - may contain errors
Lab products found in correlation
P erk erk, by Cell Signaling Technology (13 mentions)
P p38 p38, by Cell Signaling Technology (12 mentions)
P iκbα iκbα, by Cell Signaling Technology (3 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Santa Cruz Biotechnology (3 mentions)
Cleaved caspase 3, by Cell Signaling Technology (2 mentions)
Gapdh, by Cell Signaling Technology (2 mentions)
Tgf β1, by Abcam (2 mentions)
Ripa buffer, by Beyotime (2 mentions)
β actin, by Proteintech (2 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (2 mentions)
Antibodies against p iκbα, by Cell Signaling Technology (2 mentions)
Alanine transaminase, by Nanjing Jiancheng (1 mentions)
Antibody against gapdh, by Proteintech (1 mentions)
Tunel apoptosis assay kit, by Beyotime (1 mentions)
Cy3 labeled goat anti rabbit igg, by Beyotime (1 mentions)
Bcl 2, by Abcam (1 mentions)
Nfκb p nfκb, by Cell Signaling Technology (1 mentions)
P iκb, by Cell Signaling Technology (1 mentions)
Lipopolysaccharide lps from e coli 055 b5, by Merck Group (1 mentions)
Human il 6 elisa kit, by Elabscience (1 mentions)
21 protocols using p jnk jnk
1
Investigating S1P Signaling in Liver Fibrosis
2
Western Blot Analysis of MAPK and NFκB Signaling
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BMDCs (1 × 106/ml) were incubated for 30 min with 100 μg/ml of PMB with or without BHSSC (50, 100, and 150 μg/ml). Whole-cell lysates (15 μg per lane) were separated by 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and then analyzed by Western blotting using specific antibodies against ERK/p-ERK, p38/p-p38, JNK/p-JNK, and NFκB/p-NFκB (Cell Signaling Technology, MA, USA). Actin (Merck-Millipore, Darmstadt, Germany) was used as a loading control. ImageJ 1.42q software (NIH Image, National Institutes of Health; online at http://rsbweb.nih.gov/ij/ ) was used to quantify the data in the image. The density of the Actin band was used to normalize differences in loading. Expression levels were compared with those of controls.
3
Dehydrozingerone Attenuates Inflammatory Response
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Dehydrozingerone (> 99% purity) bought from TCI chemicals and structure of the compound was shown in Fig. S1A, Lipopolysaccharide (LPS from E. Coli 055: B5) purchased from Sigma Aldrich (USA). Dexamethasone was purchased from Sigma-Aldrich, USA. C-DNA synthesis kit, SYBR green mix purchased from Takara bioscience INDIA. Antibodies GAPDH, JNK P-JNK, c-JUN, p-NF-κB, NF-κB, p38, pP38, Histone-H3, IκB, p-IκB, MPO, Neutrophil Elastase were purchased from Cell signalling technology (USA), ELISA kits (IL-6, CCL2, IL-10, INF-γ, Human-IL-8 and IL-1β were bought from R&D systems. Human IL-6 ELISA kit was procured from Elabsciences (Houston, USA). Secondary Antibodies were procured from Jackson Immuno research private limited (USA). TUNEL kit was procured from Merk Millipore.
4
Investigating Molecular Mechanisms of Apoptosis
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All inhibitors were purchased from Selleck Chemicals (Houston, TX), prepared as 10 mM stock solutions in DMSO, and further diluted with culture medium before use. Annexin V-FITC/PI apoptosis detection kit and cell cycle detection kit were purchased from KeyGen BioTECH (Nanjing, China). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), DMSO, and Hoechst 33342 were obtained from Solarbio (Beijing, China). Antibodies for Src/p-Src, Mek/p-Mek, Erk/p-Erk, PI3K/p-PI3K, PDK1/p-PDK1, AKT/p-AKT, mTOR/p-mTOR, PTEN/p-PTEN(S380/T382/383), JNK/p-JNK, c-Jun, p38/p-p38, JAK2/P-JAK2, Stat3/p-Stat3, Rb/p-Rb, CDK4, CDK6, Cyclin D1, CDK2, Cyclin E1, RAD51, Bcl-2, Bax, cleaved caspase-3, and cleaved caspase-7 were all purchased from Cell Signaling Technology (Danvers, MA, USA). Tubulin and GAPDH primary antibodies were purchased from ZSGB-Biotechnology (Beijing, China). Horseradish peroxidase (HRP)-conjugated secondary antibodies against rabbit and mouse IgG were obtained from BBI life sciences (Shanghai, China).
5
Baicalin's Protective Effects in Inflammation
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Baicalin was obtained from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China), and it was resuspended using distilled water to 40 mg/mL for the animal experiments. Antibodies against P38, Bax, Bcl-2, Caspase-3, Erk1/2, Nrf2, HO-1 and β-action were purchased from Proteintech (Wuhan, China). JNK, p-JNK and all secondary antibodies used in Western blot were obtained from Cell Signaling Technology (Beverly, MA, USA). p-P38, p-Erk1/2, NQO-1, CD3 and MPO antibodies were from ABclonal Technology (Wuhan, China). CD68 antibody was provided by Abcam (Cambridge, MA). CD4 antibody was obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Trizol, Hifair™ II 1st Strand cDNA Synthesis Super Mix and Hieff®qPCR SYBR®Green Master Mix were from Yeasen (Shanghai, China). The TUNEL apoptosis and immunohistochemistry kits were obtained from Wuhan Gugeshengwu Technology Co., Ltd. (Wuhan, China). All other regular reagents were obtained from Wuhan Gugeshengwu Technology Co., Ltd. unless otherwise specified.
6
Ligularia pleurocaulis Extract Modulates Inflammatory Response
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Chemicals and antibodies EPS (Figure S1A) was isolated from the roots and rhizomes of Ligularia pleurocaulis as described previously (Yang et al., 2011) with the purity ≥98% (assessed by HPLC).
Prednisone acetate (PDN, purity >98%) (Goulding 2004 ) was obtained from Zhejiang Xianju Pharmaceutical Co., Ltd. Lipopolysaccharide (LPS) was purchased from Sigma. Rabbit mAb, mouse mAb, antibody against β-actin, p38/p-p38, ERK/p-ERK and JNK/p-JNK were purchased from Cell Signaling Technology Inc. Antibody against TLR4 and iNOS were obtained from Wanleibio Co., Ltd. Antibodies against myD88, p65/p-p65, IκBα and arginase-1 were purchased from Bioworld Technology.
All the other chemicals used in the experiments were commercial products of reagent grade.
Cell Culture and treatment RAW 264.7 cell line was obtained from the Cell Bank of Chinese Academic of Sciences. The cells were cultured in DMEM medium, supplemented with 10% fetal bovine serum and antibiotics at 37 °C in a 5% CO 2 incubator. RAW264.7 cells were pretreated with various concentrations of EPS (100, 50, 25, 12.5 μM) for 1h and then were stimulated with LPS (5 μg/mL) for 20h. Cells that were not given any treatment were used as a control.
Prednisone acetate (PDN, purity >98%) (Goulding 2004 ) was obtained from Zhejiang Xianju Pharmaceutical Co., Ltd. Lipopolysaccharide (LPS) was purchased from Sigma. Rabbit mAb, mouse mAb, antibody against β-actin, p38/p-p38, ERK/p-ERK and JNK/p-JNK were purchased from Cell Signaling Technology Inc. Antibody against TLR4 and iNOS were obtained from Wanleibio Co., Ltd. Antibodies against myD88, p65/p-p65, IκBα and arginase-1 were purchased from Bioworld Technology.
All the other chemicals used in the experiments were commercial products of reagent grade.
Cell Culture and treatment RAW 264.7 cell line was obtained from the Cell Bank of Chinese Academic of Sciences. The cells were cultured in DMEM medium, supplemented with 10% fetal bovine serum and antibiotics at 37 °C in a 5% CO 2 incubator. RAW264.7 cells were pretreated with various concentrations of EPS (100, 50, 25, 12.5 μM) for 1h and then were stimulated with LPS (5 μg/mL) for 20h. Cells that were not given any treatment were used as a control.
7
Examining Fibroblast Responses to Injury
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Tissue samples of the posterior joint capsule collected at 0, 1, 3, 7, and 14 days following injury or primary joint capsule fibroblasts treated with various drugs (MIF, TGF-β1, siRNA or inhibitors) were lysed in RIPA buffer (Beyotime, Shanghai, China) supplemented with 1 mM PMSF (Beyotime). Protein concentration in the supernatant was quantified using the Protein Assay Kit II (Bio-Rad, Inc., California, USA) to ensure equal loading. Western blots were performed as described previously 21 (link). The relative intensities of protein bands were normalized to those of β-actin. The following antibodies were used: β-actin (ProteinTech, Wuhan, China, 1:5000), MIF (Abcam, 1:1000), TGF-β1 (Abcam, 1:1000), α-smooth muscle actin (α-SMA, Abcam, 1:1000), Collagen I (Abcam, 1:1000), CD74 (Santa Cruz, CA, 1:100), p-ERK/ERK (Cell Signaling Technology, Danvers, MA, 1:1000), p-P38/P38 (Cell Signaling Technology, 1:1000), p-JNK/JNK (Cell Signaling Technology, 1:1000). Secondary antibodies included Goat anti-Rabbit IgG (H+L) (DyLight 800 4X PEG) (Invitrogen, 1:20000) and Goat anti-Mouse IgG (H+L) (DyLight 680) (Invitrogen, 1:20000). The fluorescent signals were determined with an Odyssey imaging system (Li-Cor, Lincoln, NE, USA).
8
Western Blot Analysis of Protein Expression
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Tissue samples and cells were homogenized with RIPA lysis buffer (Beyotime Biotech, Shanghai, China). Protein concentrations were determined by using a BCA protein assay kit. The proteins were separated on a 12% SDS‐polyacrylamide gel and transferred to a PVDF membrane. After blocking with 5% bovine serum albumin (BSA) in TBS, the membrane was incubated with primary antibody at 4°C overnight. The secondary antibody was conjugated to horseradish peroxidase and incubated at room temperature for 1 hour. Immunoreactivity was detected with an enhanced chemiluminescence kit (Beyotime Biotech) by a ChemiDoc™ MP System (Bio‐Rad). The following antibodies were used: HMGB1 (1:5000, Abcam), RAGE (1:1000, Abcam), RAGE (1:200, Santa Cruz), TLR4 (1:1000, Protein Tech), TLR4 (1:200, Santa Cruz) and β‐actin (1:1000, Abcam); pNF‐κB/NF‐κB, pJNK/JNK and pERK/ERK were all purchased from Cell Signaling Technology and incubated at a dilution of 1:1000.
9
Western Blot Analysis of Adipogenic and Stress Markers
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Tissue homogenate or cells lysate in lysis buffer containing 10% SDS and 1 M Tris–HCl (pH6.8) supplemented with a cocktail of protease inhibitors (cOmplete Tablets) and phosphatase inhibitors (Roche, Germany). Lysates were then quantitated and equal amounts of protein were subjected to SDS‐PAGE and immunoblotted with antibodies against HSP 90, SCD1, p‐P65/P65, p‐P38/P38, p‐JNK/JNK, PPAR γ, C/EBP α, C/EBP β, FASN, ACC. Antibodies against HSP 90, p‐P65/P65, p‐P38/P38, p‐JNK/JNK, and PPAR γ were from Cell Signaling Technology (Beverly, MA, USA), antibodies against C/EBP α was from Santa Cruz Biotechnology (Santa Cruz, CA, USA), antibodies against SCD1, FASN, and ACC were from proteintech (Wuhan, China), and antibodies against C/EBP β was from the Department of Biological Chemistry, Johns Hopkins University School of Medicine.
10
TDD Inhibits Inflammation Signaling
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TDD was synthesized in our laboratory as previously reported (Supplementary Material online, Figure S1 , 99% purity).14 LPS was purchased from Sigma‐Aldrich (St. Louis, MO, USA). Antibodies for phosphor (p)‐ERK1/2, ERK1/2, p‐p38, p38, p‐JNK, JNK, ICAM‐1, VCAM‐1, E‐selectin and NF‐κB/p65 were purchased from Cell Signaling Technology (Beverly, MA, USA). TRIzol reagent, a PrimeScript™ RT reagent kit and an SYBR Premix Ex Taq™ II kit were obtained from TaKaRa Bio, Inc. (Shiga, Japan). Lamin B Rabbit Monoclonal Antibody, a nuclear protein extraction kit, and an NF‐κB Activation‐Nuclear Translocation Assay Kit were obtained from Biotechnology Co. (Shanghai, China). The antibody for HMBOX1 was obtained from Proteintech Group Inc. (Wuhan, China). ELISA kits for IL‐6 and TNF‐α, β‐actin antibody, horseradish peroxidase‐conjugated secondary antibodies and enhanced chemiluminescence (ECL) detection kit were obtained from Boster Biotech Co., Ltd. (Wuhan, China).
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