RAW 264.7 macrophages (1 × 106 cells/mL) were cultured with SZF at concentrations of 25, 50 and 100 μg/mL, respectively. After 24 h incubation, a Trizol assay kit was applied on the conditioned cells to extract total RNA. Briefly, 1 μg of RNA was determined using qRT-PCR, and PCR amplification was carried out via incorporation of SYBR green (Roche). The amplification primer sequences GAPDH, iNOS, and COX-2 involved in this experiment were purchased from Sangon Biotech (Shanghai), GAPDH: Forward-GGTGAAGGTCGGTGTGAACG, Reverse-CTCG-CTCCTGGAAGATGGTG; iNOS: Forward-CCTCCTCGTTCAGCTCACCT, Reverse-CAATCCACAACTCGCTCCAA; COX-2: Forward- TGAGTACCGCAAA-CGCTTCT, Reverse- ACGAGGTTTTTCCACCAGCA. The expression of genes was analyzed using the 2-△△Ct method.
Cox 2
Manufactured by Sangon
Sourced in China
COX-2 is a laboratory equipment product designed for the detection and measurement of cyclooxygenase-2 (COX-2) enzyme activity. COX-2 is an important enzyme involved in the production of prostaglandins, which play a role in inflammation and other physiological processes. The COX-2 product provides a tool for researchers to study the regulation and function of this enzyme in various experimental models and biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Il 1β, by Sangon (4 mentions)
β actin, by Sangon (3 mentions)
Tnf α, by Sangon (3 mentions)
Interleukin 6 (il 6), by Sangon (3 mentions)
Sybr green, by Roche (1 mentions)
Gapdh, by Sangon (1 mentions)
Mmp 9, by Sangon (1 mentions)
Mmp 1, by Sangon (1 mentions)
M per mammalian protein extraction reagent kit, by Thermo Fisher Scientific (1 mentions)
Anti erk, by BD (1 mentions)
Anti jnk, by BD (1 mentions)
Anti p38, by BD (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Ne per nuclear and cytoplasmic extraction kit, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Actin β polyclonal antibody, by Immunoway (1 mentions)
Lps l2880, by Merck Group (1 mentions)
Imagequant software, by GE Healthcare (1 mentions)
6 protocols using cox 2
1
Quantifying Inflammatory Gene Expression
2
Antarctic Diatom Compound Evaluation
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The Antarctic diatom (Phaeodactylum tricornutum ICE-H) used in this experiment was isolated and purified from seawater and sea ice near the Zhongshan Research Station of Antarctica (S 69°48′, E 77°48′) during China’s 18th Antarctic Scientific Expedition [54 (link)]. Phaeodactylum tricornutum ICE-H was cultivated in a low-temperature light incubator after being infected in a triangle flask with f/2 media [55 (link)] at a 30% inoculation level. The light cycle was 12 L:12 D, the light intensity was 40 μmol m−2 s−1, and the culture temperature was 4 ± 1 °C. The reagent kits for the measurement of ROS, MDA, Hyp, ABTS, and DPPH were purchased from Servicebio Technology Co., Ltd. (Wuhan, China). SPF male Kunming (KM) mice were purchased from Pengyue Experimental Animal Breeding Co., Ltd. (Jinan, China). The primers targeting mRNAs, including IL-1β, TNF-α, NF-κB, COX-2, MMP-1, MMP-9, and β-actin, were synthesized by Sangon Biotech Co., Ltd. (Shanghai, China).
3
LPS-Induced Inflammatory Signaling Pathways
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Trizol, Dulbecco's modified Eagle's medium (DMEM), and other cell-culture reagents including fetal bovine serum (FBS) were obtained from Invitrogen-Gibco (Grand Island, NY, USA). Dimethyl sulfoxide (DMSO), phosphate buffered saline (PBS), Lipopolysaccharide, Escherichia coli 0127:138 (LPS), and 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-NF-κB, anti-JNK, anti-phosphorylated JNK, anti-ERK, anti-phosphorylated ERK, anti-p38, and anti-phosphorylated p38 mouse or rabbit antibodies were purchased from BD Biosciences (San Diego, CA, USA). The polymerase chain reaction (PCR) primers of iNOS, COX-2, IL-1β, IL-6, and β-actin were synthesized by Sangon Biotech Co., Ltd. (Shanghai, China). Reverse transcription and real-time PCR kits were purchased from Bio-Rad Laboratories (Hercules, CA, USA). M-PER Mammalian Protein Extraction Reagent kit and NE-PER Nuclear and Cytoplasmic Extraction kit (Thermo Scientific) were purchased from Thermo Scientific (Waltham, MA, USA).
4
Antioxidant and Anti-inflammatory Effects
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PTER (SP9570, purity ≥98%) was purchased from Beijing Solarbio Science and Technology Co., Ltd. (Beijing, China). LPS (L2880, purity >97%) was purchased from the Sigma Chemical Co. (St. Louis, MO, United States). MPO (A044-1–1), SOD (A001-3–2), CAT (A007-1–1), GSH-Px (A005-1–2) and MDA (A003-1–2) Assay Kit and were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). TNF-α, IL-6, IL-1β, COX-2 and iNOS primers were purchased from Sangon Biotech Co., Ltd. (Shanghai, China). p-p65 rabbit polyclonal antibody (A13599) was acquired from Boster Biological Technology co.Itd. p65 rabbit polyclonal antibody (BS9879M), p-IκB rabbit polyclonal antibody (BS4105), IκB rabbit polyclonal antibody (BS3601), Nrf2 rabbit polyclonal antibody (BS1258) and HO-1 rabbit polyclonal antibody (BS6626) were acquired from Bioworld Technology, Inc (Minnesota, United States). Actin-β polyclonal antibody (YT0099) was acquired from ImmunoWay Biotechnology Company (Plano, TX, United States). All other chemicals were at the reagent level.
5
Western Blot Analysis of Inflammation Markers
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The supernatant was separated from cell lysate via centrifugation at 12,000×g for 15 min at 4°C. Protein samples were separated by12% SDS-PAGE and transferred to a polyvinylidene fluoride membrane (PVDF, Millipore, Bedford, MA USA). Membranes were blocked in TBST buffer (20 mM Tris-HCl, pH 8.0, 100 mM NaCl, and 0.1% Tween-20) with 5% nonfat dry milk for 1 h. The blots were then incubated with primary antibodies anti-TLR4, NF-κB, COX2, IL-1β, TNF-α, β-actin, phospho-TLR4, and phospho-NF-κB p65 (Ser529) (Sangon, Shanghai, China) overnight at 4°C. The blots were rinsed with TBST buffer and incubated with HRP-conjugated anti-rabbit and anti-mouse secondary antibodies (at 1 : 5000 dilution, Sangon, Shanghai, China). Target proteins were visualized using chemiluminescence horseradish peroxidase (Millipore, Bedford, MA USA) and analyzed by densitometry using ImageQuant software (Molecular Dynamics, Sunnyvale, CA, USA).
6
Comprehensive Evaluation of Colitis and Inflammation
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DL (lot no: Q-016-170816) was purchased from Chengdu Ruifensi Biotechnology Co., Ltd. (Chengdu, China) (Figure 1 ).The dextran sulfate sodium (DSS) (lot no. Q1723) was purchased from USA MP Ltd. Mesalazine SR Granules (lot no. 160627) was purchased from Shanghai Aizhifa Pharmaceutical Co., Ltd. (Shanghai, China). Stool blood test kit was purchased from Zhuhai Besso Biotechnology Co., Ltd. (Zhuhai, China). TNF-α, IL-1β, MCP-1, MPO, SOD, IL-6, IL-17, and IL-23 ELISA assay were purchased from Beijing Baiolaibo Technology Co., Ltd. (Beijing, China). iNOS, COX2, IL-6, GP130, STAT3, IL-17, IL-23, MUC2, XBP1s, and α-defensins (polymerase chain reaction RT-PCR) were purchased from Shanghai Sangon Biotech (Shanghai, China).
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