The levels of inflammation factors in heart tissue or H9C2 cells was measured using IL-1β (Elabscience, China), IL-6 (Elabscience, China) and TNF-α (Elabscience, China) ELISA kits according to the manufacturer’s recommendations, The OD is measured spectrophotometrically at a wavelength of 450 nm, the contents of IL-1β, IL-6 and TNF-α were calculated according to the standard curve.
Tnf α
Manufactured by Elabscience
Sourced in China, United States
Tumor Necrosis Factor-alpha (TNF-α) is a cytokine that plays a crucial role in the inflammatory response and immune system regulation. It is involved in a wide range of biological processes, including cell proliferation, differentiation, and apoptosis.
Automatically generated - may contain errors
Lab products found in correlation
Interleukin 6 (il 6), by Elabscience (21 mentions)
Il 1β, by Elabscience (19 mentions)
Il 10, by Elabscience (7 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (3 mentions)
Mmp 13, by Elabscience (2 mentions)
8 oh dg, by Elabscience (2 mentions)
Caspase 3, by Elabscience (2 mentions)
Tgf β, by Elabscience (2 mentions)
Gapdh, by Cell Signaling Technology (2 mentions)
Nf κb, by Cell Signaling Technology (2 mentions)
Ifn γ, by Elabscience (2 mentions)
Hmgb 1, by Elabscience (2 mentions)
Interleukin 4 (il 4), by Elabscience (2 mentions)
Il 12, by Elabscience (1 mentions)
Mmp 3, by Elabscience (1 mentions)
Mmp 1, by Elabscience (1 mentions)
F4 80, by Cell Signaling Technology (1 mentions)
P stat5, by Cell Signaling Technology (1 mentions)
Stat5, by Cell Signaling Technology (1 mentions)
P nf κb, by Cell Signaling Technology (1 mentions)
56 protocols using tnf α
1
Inflammatory Cytokine Quantification
2
Complement System Activation After ROSC
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Venous blood samples were collected, respectively, from the patients at 2 h, 72 h and 168 h after ROSC or healthy volunteers on enrolled day. We have collected the blood samples at 2 h of all patients who survived to 1 day or longer, the blood samples at 72 h only in patients who survived to 3 days or longer after ROSC, and the blood samples at 168 h only in patients who survived to 7 days or longer after ROSC. Sample collectors, clinical investigators, assistants, and laboratory personnel were unaware of the study protocol. Blood samples were centrifuged (2500×g) at 4 °C for 10 min to collect the serum. The serum samples were aliquoted and stored at − 80 °C until further analysis. Enzyme-linked immunosorbent assay (ELISA) kits were used to measure serum sCD59 (Elabscience, Wuhan, China), sC5b-9 (Elabscience, Wuhan, China), C1q (Elabscience, Wuhan, China), MBL (CUSABIO, Wuhan, China), Bb (Quidel, California, USA), C3a (CUSABIO, Wuhan, China), C3b (CUSABIO, Wuhan, China), C5a (CUSABIO, Wuhan, China), IL-6 (Elabscience, Wuhan, China), TNF-a (Elabscience, Wuhan, China), NSE (CUSABIO, Wuhan, China) and S100β (CUSABIO, Wuhan, China) in accordance with the manufactures' instructions.
3
Inflammatory Cytokine ELISA Quantification
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4
Quantification of Nitric Oxide in Plasma
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MMP-1, MMP-3, MMP-13, Collagen II, IL-1β, TNF-α, IL-6, and iNOS were purchased from Elabscience, Hubei, China. NF-κB p65, COX-2, and PGE2 were purchased from Taiclone (Taipei, Taiwan). All analyses were performed according to the manufacturer’s instructions. Nitric oxide (NO) production was analyzed by the Griess reagent method. 100 μL of plasma and serially diluted standard (0.1 M Nitrite) were added to the 96-well plate. Then, 50 μL of SUL solution (0.1% sulfanilamide solution in 2.5% phosphoric acid), and 50 μL of NED solution (0.1% N-(1-naphthyl) ethylenediamine dehydrate chloride in dd water) were added. After 10 min, the absorbance was measured at 540 nm. The amount of NO was calculated by plotting the standard curve66 .
5
Ovarian Oxidative and Inflammatory Biomarkers
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About 0.1 g of the right ovaries of each animal were homogenized on ice in 10 ml of cold phosphate buffer solution. The resultant solutions were centrifuged at 10,000 rpm for15 mins in cold centrifuge after which the supernatant was decanted into tubes and stored at −20°C for biochemical assay later.
The ovarian concentrations of UA [26 (link)], MDA [27 (link)], and GSH [28 (link)] were assayed by colorimetric method as previously reported. The activities of XO [27 (link)], superoxide dismutase (SOD) [28 (link)], catalase [28 (link)], glutathione peroxidase (GPx) [29 (link)], glutathione-S-transferase (GST) [27 (link)], and myeloperoxidase (MPO) [29 (link)] in the ovarian tissues were assayed by colorimetry as earlier documented. Also, the ovarian levels of TNF-α (Elabscience Biotechnology Inc., U.S.A.), IL-6 (Elabscience Biotechnology Inc., U.S.A.), 8OHdG (Elabscience Biotechnology Inc., U.S.A.), and VCAM-1 (Elabscience, Wuhan, China) as well as the activity of caspase 3 (Elabscience Biotechnology Inc., U.S.A.) were determined using ELISA kit following the manufacturers’ guidelines. Ovarian DNA fragmentation index was determined by diphenylamine methods as previously reported [25 ].
The ovarian concentrations of UA [26 (link)], MDA [27 (link)], and GSH [28 (link)] were assayed by colorimetric method as previously reported. The activities of XO [27 (link)], superoxide dismutase (SOD) [28 (link)], catalase [28 (link)], glutathione peroxidase (GPx) [29 (link)], glutathione-S-transferase (GST) [27 (link)], and myeloperoxidase (MPO) [29 (link)] in the ovarian tissues were assayed by colorimetry as earlier documented. Also, the ovarian levels of TNF-α (Elabscience Biotechnology Inc., U.S.A.), IL-6 (Elabscience Biotechnology Inc., U.S.A.), 8OHdG (Elabscience Biotechnology Inc., U.S.A.), and VCAM-1 (Elabscience, Wuhan, China) as well as the activity of caspase 3 (Elabscience Biotechnology Inc., U.S.A.) were determined using ELISA kit following the manufacturers’ guidelines. Ovarian DNA fragmentation index was determined by diphenylamine methods as previously reported [25 ].
6
Quantifying Kidney Cytokine Levels
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7
Quantification of Inflammatory Mediators
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The cell culture supernatants were collected and centrifuged at 1,000 g for 20 min. Concentrations of MCP-1 (Fcmacs Biotechnology Co., Ltd., Nanjing, China) and TNF-α (Elabscience Biotechnology Co., Ltd., Wuhan, China) in the supernatants were detected by ELISA kits according to the manufacturer's instructions.
8
Detailed Inflammatory Response Protocol
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SHI, with purity over 98%, was obtained from Shanghai Yuanye Bio-Technology Co., Ltd. Dexamethasone (DXM) was supplied from Xiansheng Drug Store (Nanjing, China). The drugs were dissolved by DMSO [with the concentration of DMSO less than 0.1% (w/v)]. LPS (Escherichia coli 055:B5) was produced by Sigma (St. Louis, United States). Mouse TNF-α, IL-1β, IL-6, IL-10, and TGF-β were purchased from Elabscience (Wuhan, China). The myeloperoxidase (MPO) commercial kit was provided by Jiancheng (Nanjing, China). The primary antibodies ECM1, Ly6G, F4/80, p-STAT5, STAT5, p-NF-κB, NF-κB, CD16/32, iNOS, CD206, Arg1, and GAPDH were produced by Cell Signaling Technology (CST) (Danvers, MA, United States).
9
Cytokine Quantification in Brain Tissue
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Frozen brain was powered and homogenized in lysis buffer (10% RIPA buffer, Merk Millipore, MA, USA), added 1 mM phenylmethylsulfonyl fluoride (PMSF) (Amresco, Solon, OH, USA), 10 mM sodium orthovanadate (Sigma-Aldrich), 100 mM sodium fluoride (LabSynth, Diadema, SP, Brazil), 10 mM sodium pyrophosphate (Sigma-Aldrich), and 0.2% protease inhibitor (P8340, Sigma-Aldrich), for 30 min at 4 ºC, and centrifuged (15.000x g, 4 ºC, 20 min) to obtain supernatant. Total proteins were quantified in supernatant (PierceTM BCA Protein Assay Kit, ThermoFisher Scientific), and interferon gamma (IFN-γ) (Sigma-Aldrich), IL-1β, IL-6, TNFα, (Elabscience, Houston, TX, USA) and IL-10 (Abcam, Cambridge, MA, USA) were measured by ELISA, according to the manufactures instructions. Results are presented as cytokine concentration per mg of protein.
10
Curcumin Attenuates Inflammatory Responses
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Curcumin (purity
>98.0%) was obtained from Sigma Technology (Danvers). Nissl staining
solution was purchased from Beyotime Biotechnology Co., Ltd. (Shanghai,
China). TNF-α and IL-1β enzyme-linked immunosorbent assay
(ELISA) kits were provided by Elabscience Biotechnology Co., Ltd.
(Wuhan, China). 4′,6-Diamidino-2-phenylindole (DAPI) was purchased
from Solarbio Science (Beijing, China). The ApoE4, COX-2, iNOS, NF-κBp65,
p-NF-κBp65, GRP78, IRE1α, β-actin, and IgG primary
antibodies and Alexa Fluor 488 fluorescent secondary antibodies were
all purchased from Abcam and were used at the indicated dilutions
for western blotting and immunostaining.
>98.0%) was obtained from Sigma Technology (Danvers). Nissl staining
solution was purchased from Beyotime Biotechnology Co., Ltd. (Shanghai,
China). TNF-α and IL-1β enzyme-linked immunosorbent assay
(ELISA) kits were provided by Elabscience Biotechnology Co., Ltd.
(Wuhan, China). 4′,6-Diamidino-2-phenylindole (DAPI) was purchased
from Solarbio Science (Beijing, China). The ApoE4, COX-2, iNOS, NF-κBp65,
p-NF-κBp65, GRP78, IRE1α, β-actin, and IgG primary
antibodies and Alexa Fluor 488 fluorescent secondary antibodies were
all purchased from Abcam and were used at the indicated dilutions
for western blotting and immunostaining.
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