For myoblast proliferation experiments, growing myoblasts (70–80% confluence) were transfected with pEGFP-N1-Akirin1 or the pEGFP-N1 empty plasmids using Lipofectin3000 (Invitrogen, U.S.A.) according to the manufacturer’s instructions. At 24 h post-transfection, cells were harvested for RNA and protein extraction. For myoblast differentiation experiments, pEGFP-N1-Akirin1 plasmids or the pEGFP-N1 plasmids were transfected for 24 h before the induction of differentiation. At 12, 24, 36, and 48 h after the induction of differentiation, cells were harvested for RNA and protein extraction, respectively. For LY294002 or SB203580 treatment experiments, pEGFP-N1-Akirin1 plasmids or the pEGFP-N1 plasmids were transfected for 24 h before the induction of differentiation, when GM was switched to DM, LY294002 (20 μM, PI3K inhibitor) (Beyotime Biotech, China) or SB203580 (10 μM, p38 MAPK inhibitor) (Beyotime Biotech, China) were added into the duck myoblasts [16 (link)]. After LY294002 or SB203580 treatment, cells were harvested for RNA and protein extraction, respectively.
Sb203580
Manufactured by Beyotime
Sourced in China, United States
SB203580 is a selective inhibitor of p38 mitogen-activated protein kinase (MAPK). It is a small molecule that binds to and inhibits the enzymatic activity of the p38 MAPK isoforms.
Automatically generated - may contain errors
Lab products found in correlation
Sp600125, by Beyotime (31 mentions)
U0126, by Beyotime (17 mentions)
Bay 11 7082, by Beyotime (11 mentions)
Ly294002, by Beyotime (8 mentions)
Pd98059, by Beyotime (8 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (5 mentions)
β actin, by Santa Cruz Biotechnology (4 mentions)
Dimethyl sulfoxide (dmso), by Beyotime (3 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (3 mentions)
Luminoskan ascent, by Thermo Fisher Scientific (3 mentions)
Ly294002, by Merck Group (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
M csf, by Thermo Fisher Scientific (2 mentions)
Anti nf κb p65, by BioLegend (2 mentions)
Hrp conjugated anti mouse and anti goat igg, by Santa Cruz Biotechnology (2 mentions)
Phospho erk1 2, by BD (2 mentions)
Tp0427736 hcl, by Selleck Chemicals (2 mentions)
P jnk, by Beyotime (2 mentions)
67 protocols using sb203580
1
Akirin1 Regulates Myoblast Proliferation and Differentiation
2
Bile Acids and Kinase Inhibitor Preparation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
UA and TCDCA were purchased from Sigma Aldrich (St. Louis, MO, USA). UA was dissolved in deionized water and filtered and was free of crystals (by polarizing microscopy), the stock solution concentration was 50 mg/ml. TCDCA stock solution was prepared in sterile double distilled water at 200 mM. SB203580 was purchased from Beyotime (Shanghai, China), SB203580 stock solution was prepared in sterile double distilled water at 100 μM.
3
Signaling Pathways in H2O2-Induced Granulosa Cell Responses
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H89, LY294002, SB203580 and U0126 were purchased from Beyotime (Beijing, China). After exposure to 200 μM H2O2 (Sigma, St. Louis, MO, USA) for 1 h, MGCs were rinsed with PBS and grown in serum-free DMEM/F-12 containing 7.5 IU/ml FSH for 2, 6, 12, 24 or 36 h. In some experiments, H89 (10 μM), LY294002 (20 μM), SB203580 (20 μM) or U0126 (3 μM) was added 30 min before FSH treatment.
4
Reagents and Antibodies for Cell Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Reagents. Neutral red staining solution was obtained from Sangon Biotech (Shanghai, China). U0126 (MEK1/2 inhibitor), SP600125 (JNK inhibitor) and SB203580 (p38 inhibitor) were purchased from Beyotime (Shanghai, China). LY294002 (PI3K inhibitor) was purchased from Selleck Chemicals (Houston, TX, USA). Anti-FIP-glu antiserum was raised in rabbits (99) . Anti-6×His Tag mouse monoclonal antibody, HRP-conjugated Goat Anti-Mouse IgG and HRP-conjugated Goat Anti-Rabbit IgG were from Sangon Biotech (Shanghai, China).
5
LPS-Induced Inflammatory Response in RAW 264.7 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RAW 264.7 cells were cultured in 6-well culture dishes until they reached approximately 80% confluence and were then stimulated with 1 μg/mL Escherichia coli LPS (InvivoGen, San Diego, CA, USA) for a certain number of hours. Cells not stimulated by LPS were used as a control.
In the signaling pathway inhibition experiments, the cells were first treated with the signaling pathway inhibitor BAY 11-7082 (Beyotime, Shanghai, China; 10 μM), U0126 (Beyotime, Shanghai, China; 10 μM), SB203580 (Beyotime, Shanghai, China; 20 μM) or SP600125 (Beyotime, Shanghai, China; 20 μM) for 1 h, and were then stimulated with 1 μg/mL LPS for 6 h. Cells not stimulated with LPS or treated with signaling pathway inhibitors were used as a blank control.
In the signaling pathway inhibition experiments, the cells were first treated with the signaling pathway inhibitor BAY 11-7082 (Beyotime, Shanghai, China; 10 μM), U0126 (Beyotime, Shanghai, China; 10 μM), SB203580 (Beyotime, Shanghai, China; 20 μM) or SP600125 (Beyotime, Shanghai, China; 20 μM) for 1 h, and were then stimulated with 1 μg/mL LPS for 6 h. Cells not stimulated with LPS or treated with signaling pathway inhibitors were used as a blank control.
6
Bovine IL-1α Signaling Pathway Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Recombinant bovine IL-1α was purchased from Kingfisher Biotech. U0126, SP600125, SB203580 and BAY11-7082 were obtained from Beyotime Biotechnology. Erk antibody, p-Erk antibody, c-Jun antibody, p-c-Jun antibody, IκBα antibody and p-IκBα antibody were purchased from Cell Signaling Technology. p38 antibody, p-p38 antibody and β-actin antibody were purchased from LifeSpan BioSciences. Forskolin were purchased from Abcam.
7
Solid-Phase Synthesis of EM-1 and EM-2 Peptides
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
EM-1 and EM-2 were prepared by manual solid-phase synthesis using standard N-fluorenylmethoxycarbonyl (Fmoc) chemistry as reported in our previous study [32 (link)]. Fmoc-protected amino acids (GL Biochem Ltd., China) were coupled with Rink amide 4-methybenzhydrylamine (MBHA) resin (Tianjin Nankai Hecheng Science & Technology Co., Ltd., China). The crude peptides were purified by preparative reversed-phase HPLC (RP-HPLC) and determined by electrospray ionization mass spectrometer (ESI-Q-TOF maXis-4G, Bruker Daltonics).
Naloxone, β-FNA, nor-binaltorphimine (nor-BNI), and naltrindole (NTI) were obtained from Sigma-Aldrich. The selective p38 MAPK inhibitor SB203580 was purchased from Beyotime Institute of Biotechnology and dissolved in 1% DMSO in saline. All other drugs were dissolved in sterilized saline and stored at − 20 °C.
Naloxone, β-FNA, nor-binaltorphimine (nor-BNI), and naltrindole (NTI) were obtained from Sigma-Aldrich. The selective p38 MAPK inhibitor SB203580 was purchased from Beyotime Institute of Biotechnology and dissolved in 1% DMSO in saline. All other drugs were dissolved in sterilized saline and stored at − 20 °C.
8
Evaluating RPMTG's Effects on CRC Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human CRC cell lines HCT116 and SW620 were purchased from The Cell Bank of Type Culture Collection of The Chinese Academy of Sciences. HCT116 and SW620 cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco; Thermo Fisher Scientific, Inc.), supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 100 µg/ml streptomycin. The HCT116 (1×104 cells/well) and SW620 (2×104 cells/well) cells were pretreated with JNK inhibitor (SP600125, 10 µM; Beyotime Institute of Biotechnology) and p38 inhibitor (SB203580, 10 µM; Beyotime Institute of Biotechnology) for 2 h, and then treated with 250 µg/ml RPMTG for 24 h in a 37°C incubator containing 5% CO2. In subsequent experiments, the effects of RPMTG combined with JNK inhibitors or p38 inhibitors on proliferation and apoptosis were evaluated.
9
Cobrotoxin-Induced Cytotoxicity Mechanism
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cobrotoxin was a gift from the Department of Pharmacy at the Suzhou University Medical College (Suzhou, Jiangsu, China), while the 3-methyl adenine (3-MA), dimethyl sulfoxide (DMSO) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). SB203580 was purchased from Beyotime Institute of Biotechnology (Haimen, China) and an automatic enzyme standard instrument (Benchmark) was purchased from Bio-Rad (Hercules, CA, USA). A Heracell 150 carbon dioxide incubation box was purchased from Thermo Fisher Scientific (Rockford, IL, USA). SDS-PAGE apparatus (Mini-PROTEAN Tetra Electrophoresis System and Mini-PROTEAN 3 Dodeca Cell, Bio-Rad) and a FEI TECNAI 10 transmission electron microscope (TEM; Philips, Amsterdam, Holland) were also utilized.
10
Hypoxia-Induced HaCaT Cell Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HaCaT cells were purchased from the cell bank of the Chinese Academy of Sciences in Beijing, China. Cells were cultured in RPMI 1640 medium (HyClone, USA) containing 10% fetal bovine serum (HyClone, USA), 100 U/ml penicillin (Invitrogen, USA), and 100 mg/ml streptomycin (Invitrogen, USA).The cells were then put in 37°C, 5% CO2 incubator, and 95% humidity.
Hypoxic conditions of 1% O2, 5% CO2, and 94% N2 were produced utilizing an oxygen control incubator (model: 3131; Thermo Scientific). The p38/MAPK inhibitor SB203580 (Beyotime) (5 μmol/l) was added to these cultures and incubated at 37°C for 30 minutes before hypoxia treatment. In addition the ADAM17 inhibitor, TAPI-2 (40 μM) was added to the cultures and incubated at 37°C for 12 hours before hypoxia treatment.
Hypoxic conditions of 1% O2, 5% CO2, and 94% N2 were produced utilizing an oxygen control incubator (model: 3131; Thermo Scientific). The p38/MAPK inhibitor SB203580 (Beyotime) (5 μmol/l) was added to these cultures and incubated at 37°C for 30 minutes before hypoxia treatment. In addition the ADAM17 inhibitor, TAPI-2 (40 μM) was added to the cultures and incubated at 37°C for 12 hours before hypoxia treatment.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!