All serum samples were tested for HMGB1 (FineTest, Wuhan, China), tumor necrosis factor-α (TNF-α) (Wanleibio, Shenyang, China), IL-1β (Wanleibio, Shenyang, China), and IL-6 (Wanleibio, Shenyang, China), using a rat ELISA kit. The ELISA assay was performed following the manufacturer's protocols. The optical density of each well was measured spectrophotometrically at 450 nm within 5 min after adding the stop solution using a microplate reader (ELX-800, BIOTEK, USA).
Interleukin 6 (il 6)
Manufactured by Wanlei
Sourced in China
IL-6 is a cytokine that plays a central role in immune and inflammatory responses. It is a soluble protein that acts as a signaling molecule between cells. IL-6 is involved in various biological processes, including the regulation of the immune system, inflammation, and hematopoiesis.
Automatically generated - may contain errors
Lab products found in correlation
Tnf α, by Wanlei (3 mentions)
Il 1β, by Wanlei (2 mentions)
Elx800, by Agilent Technologies (1 mentions)
E cadherin, by Proteintech (1 mentions)
N cadherin, by Proteintech (1 mentions)
Vimentin, by Proteintech (1 mentions)
Nf κb, by Wanlei (1 mentions)
Tnfsf9, by Proteintech (1 mentions)
Snail, by Wanlei (1 mentions)
β catenin, by Proteintech (1 mentions)
Gapdh, by Proteintech (1 mentions)
P jak2, by Affinity Biosciences (1 mentions)
Mpm 2, by Abcam (1 mentions)
Ki 67, by Wanlei (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
Cell cycle analysis kit, by Beyotime (1 mentions)
β actin, by Affinity Biosciences (1 mentions)
Gapdh, by Affinity Biosciences (1 mentions)
Cell counting kit 8, by Beyotime (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
6 protocols using interleukin 6 (il 6)
1
Serum Biomarker Assessment in Rat Model
2
Protein Expression Profiling in Cell Signaling
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First, the total cell protein was extracted with protein extract (Beyotime, China). Then, 10% sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE) was used for electrophoresis, and the protein was transferred to polyvinylidene (PVDF) and sealed with 5% skimmed milk for 1 h. It was then incubated overnight at 4°C with the following primary antibodies: GAPDH (1:10000, ProteinTech), TNFSF9 (1:2000, ProteinTech), β-catenin (1:1000, ProteinTech), Wnt (1:1000, Wanleibio), ERK (1:500, Wanleibio), phosphorylated-ERK (1:1000, Santa), JNK (1:1000, Wanleibio), phosphorylated-JNK (1:1000, Wanleibio), AKT (1:1000, Santa), phosphorylated-AKT (1:1000, Santa), NF-ΚB (1:1000, Wanleibio), phosphorylated-NF-ΚB (1:1000, Wanleibio), Snail (1:1500, Wanleibio), focal adhesion kinase (FAK) (1:2000, ProteinTech), proto-oncogene tyrosine-protein kinase Src (Src) (1:800, ProteinTech), IL-6 (1:5000, Wanleibio), IL-8 (1:1000, Wanleibio), TNF-α (1:1000, Santa), E-cadherin (ProteinTech, 1:10000), N-cadherin (ProteinTech, 1:3000), and Vimentin (ProteinTech, 1:2000). Finally, the membrane was incubated with appropriate secondary antibodies for 1 h at room temperature. The band was quantitatively measured using the Zeiss LSM800 spectrometer (Carl Zeiss AG, Germany) [44 (link)].
3
Serum Biomarker Quantification in Mice
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The enzyme‐linked immunosorbent assay (ELISA) kits were used to detect the expression level of IL‐6 (Wanleibio, China), IL‐1β (Wanleibio, Shenyang, China), TNF‐α (Wanleibio, Shenyang, China), Lactic dehydrogenase (LDH; Wanleibio, Shenyang, China), and total protein (TP; Nanjing Jiancheng Bioengineering Institute, China) in mice serum according to the manufacturer instructions.
4
Gastric Cancer Cell Lines: Culture and Evaluation
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Human gastric cancer cell lines HGC-27 and AGS were purchased from Procell Life Science and Technology Ltd. Gastric cancer cells HGC-27 and AGS were cultured in Dulbecco's modified Eagle's medium (DMEM) and RPMI 1640, respectively, supplemented with 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin solution in a 37°C cell incubator with 5% CO2.
DMEM and RPMI 1640 medium were purchased from HyClone (USA). TAS-102, FTD and CTS were obtained from TargetMol (China), and their purity was greater than 98%. Cell Counting Kit (CCK)-8 and cell cycle analysis kit were obtained from Beyotime (China), and Annexin V-FITC/PI double staining cell apoptosis detection kit was purchased from KeyGEN BioTHCE (China). Primary antibodies included BrdU, Cyclin B1, Cyclin D1, Caspase 3, Caspase 9, Bcl-2, Survivin, phospho-extracellular signal-regulated kinase (p-ERK) (Thr202/Tyr204) and ERK, p-AKT (Ser473) and AKT, p-STAT3 (Tyr705) and STAT3 (Cell Signaling Technology), p-JAK2 (Tyr221) and JAK2, GAPDH, β-actin (Affinity), MPM-2 (Abcam), IL-6 and Ki-67 (Wanleibio). Secondary antibodies horseradish peroxidase (HRP)-conjugated goat anti-rabbit/mouse IgG and Cy3 labeled goat anti-mouse IgG were from ZSGB-Bio (China) and Abclonal (China), respectively.
DMEM and RPMI 1640 medium were purchased from HyClone (USA). TAS-102, FTD and CTS were obtained from TargetMol (China), and their purity was greater than 98%. Cell Counting Kit (CCK)-8 and cell cycle analysis kit were obtained from Beyotime (China), and Annexin V-FITC/PI double staining cell apoptosis detection kit was purchased from KeyGEN BioTHCE (China). Primary antibodies included BrdU, Cyclin B1, Cyclin D1, Caspase 3, Caspase 9, Bcl-2, Survivin, phospho-extracellular signal-regulated kinase (p-ERK) (Thr202/Tyr204) and ERK, p-AKT (Ser473) and AKT, p-STAT3 (Tyr705) and STAT3 (Cell Signaling Technology), p-JAK2 (Tyr221) and JAK2, GAPDH, β-actin (Affinity), MPM-2 (Abcam), IL-6 and Ki-67 (Wanleibio). Secondary antibodies horseradish peroxidase (HRP)-conjugated goat anti-rabbit/mouse IgG and Cy3 labeled goat anti-mouse IgG were from ZSGB-Bio (China) and Abclonal (China), respectively.
5
Protein Expression Profiling in Tissue Lysates
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RIPA buffer containing protease inhibitors (Beyotime) was used to lyse tissues. The primary antibodies were as follows: IL-6 (Wanleibio, Shenyang, China,), TNF-α (Wanleibio), Caspase-3 (Cell Signaling Technology, Danvers, MA, USA), iNOS (Santa Cruz Biotechnology, Santa Cruz, CA, USA), Bax (Cell Signaling Technology), SIRT 1 (Wanleibio), p53 (Wanleibio), p21 (Wanleibio), SA-β-gal (Cell Signaling Technology), p16 (Wanleibio), Nrf2 (Beyotime), HO-1 (Wanleibio), and β-actin (Wanleibio). The secondary antibodies were as follows: HRP-labeled goat anti-mouse and anti-rabbit IgG(H + L) (Beyotime).
6
Western Blotting for Hippocampal Protein Analysis
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The protocol of western blotting (WB) was performed as previously described (Wei et al., 2022 (link)). In brief, the hippocampal tissue was lysed in RIPA lysis buffer (Beyotime, P0013B) and centrifuged at 12,000 × g for 15 min to collect the supernatant. The protein samples were electrophoretically separated and then blotted onto polyvinylidene fluoride membranes (Millipore, IPVH00010). The protein levels were determined via incubation with antibodies against PGC‐1α (1:1000; abcam), SIRT1 (1:500; Santa Cruz), NF‐κB (1:5000; abcam), brain‐derived neurotrophic factor (BDNF; 1:1000; abcam), PSD‐95 (1:2000; abcam), SYN (1:1000; bioss), IL‐1β (1:500; bioss), IL‐6 (1:1000; Wanleibio), and TNF‐α (1:1000; bioss). The membranes were then incubated with horseradish peroxidase‐labeled secondary antibodies (1:20,000; Zsbio) according to the properties of the primary antibody. Protein detection was performed using an ECL luminescence kit (Thermo, 340958) and quantified with ImageJ software.
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