The buds of Castanopsis lamontii were collected in the Dehong Area, Mang City, Yunnan Province, China (24°17′12″ N, 98°23′6″ E) and identified by Botanist Keke Chen, Kunming Institute of Botany, Chinese Academy of Sciences. The hydroxyl free radical assay kit and superoxide anion assay kit were obtained from Nanjing Jiancheng Bioengineering Institute. The LDH cytotoxicity assay kit, NO assay kit, primary antibodies (p-NF-κB (p65) (Product #AN371), NF-κB (Product #AF0246), GAPDH (Product #AF0006), and secondary antibodies (Product #A0216 & A0208) were purchased from Beyotime Institute of Biotechnology, Co. Ltd. (Haimen, China) Primary antibodies (TLR4 (Product #14358S), COX-2 (Product #12282), iNOS (13120S) were purchased from Cell Signaling Technology, Co. Ltd. (Danvers, MA, USA). The MTT cell proliferation and cytotoxicity assay kit was purchased from Nanjing KeyGEN Bio-TECH, Co. Ltd. (Nanjing, China) The PGE2 ELISA kit and the TNF-α ELISA kit were purchased from Jiangsu Meimian industrial Co., Ltd. (Zhangjiagang, China) Chemical standards used in HPLC analysis, UPLC-Q-TOF/MS analysis and cell assays were purchased from Sigma-Aldrich Co., Ltd. (St. Louis, MO, USA).
Tnf α elisa kit
Manufactured by Jiangsu Meimian
Sourced in China
The TNF-α ELISA kit is a laboratory equipment used for quantitative measurement of tumor necrosis factor alpha (TNF-α) in biological samples. It is a sandwich enzyme-linked immunosorbent assay (ELISA) that employs the quantitative sandwich enzyme immunoassay technique.
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Lab products found in correlation
Il 6 elisa kit, by Jiangsu Meimian (18 mentions)
Il 1β elisa kit, by Jiangsu Meimian (9 mentions)
Superoxide anion assay kit, by Nanjing Jiancheng (3 mentions)
Hydroxyl free radical assay kit, by Nanjing Jiancheng (3 mentions)
Nf κb, by Beyotime (3 mentions)
Ldh cytotoxicity assay kit, by Beyotime (3 mentions)
Gapdh, by Beyotime (3 mentions)
Mtt cell proliferation and cytotoxicity assay kit, by Keygen Biotech (3 mentions)
Collagenase type 1, by Merck Group (1 mentions)
Membrane and cytosol protein extraction kit, by Beyotime (1 mentions)
Anti cd31 antibody, by Proteintech (1 mentions)
Calcein am pi kit, by Solarbio (1 mentions)
N cadherin antibody, by Proteintech (1 mentions)
Epcam antibody, by Proteintech (1 mentions)
Rat tail tendon collagen type 1, by Solarbio (1 mentions)
Dcfh da, by Beyotime (1 mentions)
Interleukin 6 (il 6), by Jiangsu Meimian (1 mentions)
Il 1β, by Jiangsu Meimian (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by Beyotime (1 mentions)
6 protocols using tnf α elisa kit
1
Bioactive Components of Castanopsis lamontii
2
Serum TNF-α and IL-6 Quantification
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ELISA was performed according to the Mouse Tumor Necrosis Factor α (TNF-α) ELISA Kit and Mouse Interleukin-6 (IL-6) ELISA Kit instructions (Jiangsu Meimian Industrial Co., Ltd., Yancheng, China). Serum levels of TNF-α and IL-6 were determined at OD 450 nm and corresponded to standard curves.
3
Quantification of Inflammatory Cytokines
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And the concentrations of IL-6, IL-1β, TNF-α in cell supernatants were measured using bovine IL-6 ELISA kit (MM-34736O1, Meimian, Jiangsu), IL-1β ELISA kit (MM-36949O1, Meimian, Jiangsu), TNF-α ELISA kit (MM-1568O1, Meimian, Jiangsu) according to the manufacturer's protocols.
4
Cytokine Secretion in Chondrocytes
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The chondrocytes were seeded into 6-well culture plates in dMEM/F12 + 10% FBS and incubated for 24 h. Following pre-treatment with HT, the cells were exposed to AOPPs for 24 h. Following this, the cell supernatants were collected and centrifuged at 12,000 x g for 15 min at 4˚C. The TNF-α ELISA kit (MEIMIAN) and IL-6 ELISA kit (MEIMIAN) were then used to detect cytokine secretion into the supernatant. The Od values were determined at 450 nm using a spectrophotometric plate reader (Molecular devices, LLc). All experiments were performed in triplicate.
Statistical analysis. Data are presented as the mean ± SEM of three independent experiments. One-way ANOVA followed by Tukey's post hoc test was used to determine the differences between groups. P<0.05 was considered to indicate a statistically significant difference. All statistical analyses were performed using GraphPad Prism version 6.0 (GraphPad Software, Inc.).
Statistical analysis. Data are presented as the mean ± SEM of three independent experiments. One-way ANOVA followed by Tukey's post hoc test was used to determine the differences between groups. P<0.05 was considered to indicate a statistically significant difference. All statistical analyses were performed using GraphPad Prism version 6.0 (GraphPad Software, Inc.).
5
Multifunctional Nanoparticles for Cancer Therapy
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Ethylene glycol (EG), poly(vinyl pyrrolidone) (PVP), silver trifluoroacetate (CF3COOAg), doxorubicin hydrochloride (Dox), 2-iminothiolane (Traut’s reagent), MTT and iohexol were obtained from Aladdin Co., Ltd. (Shanghai, China). Sodium hydrosulfide hydrate (NaHS) was purchased from Saen Chemical Technology Co., Ltd (Shanghai, China). Chloroauric acid (HAuCl4) was obtained from Shanghai Titan Scientific Co., Ltd. (Shanghai, China). Membrane and Cytosol Protein Extraction Kit, DCFH-DA, 4′6-diamidino-2-phenylindole (DAPI) and Annexin V-FITC/PI Apoptosis Detection Kit were purchased from Beyotime (China). Collagenase (Type I) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Rattailtendon collagen type I and Calcein-AM/PI kit were purchased from Solarbio (Beijing, China). DSPE-PEG2k-MAL was purchased from ShangHai PonsureBio Tech. Inc. (Shanghai, China). SH-PEG-Cy5.5 was purchased from ShangHai ToYongBio Tech.Inc. (Shanghai, China). N-cadherin antibody, anti-CD47 antibody, EpCAM antibody, ATP1A1 antibody, collagen type I antibody and anti-CD31 antibody were purchased from Proteintech (Wuhan, China). Nab-paclitaxel was purchased from Qilu pharmaceutical (Hainan, China). IL-6, IL-1β and TNF-α ELISA kits were obtained from Jiangsu Meimian Industrial Co., Ltd. (Jiangsu, China).
6
Measuring Inflammatory Cytokines in Mice
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ELISA assay was performed to measure the serum concentrations of Ccl2, Cxcl8 and Tnf-α ELISA Kits (MEIMIAN) in mouse orthotopic syngeneic graft models according to the manufacturer’s instructions.
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