InVivoMab anti-mouse Ly6G (CAT#: BE0075-1) was purchased from Bio X Cell (Lebanon, NH, USA). Compound C (CpC, CAT#: 171260), LPS (E. coli O111:B4, CAT#: L4391), STO-609 (STO, CAT#: 570250) and lucigenin (CAT#: M8010) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Mouse interleukin-1β (IL-1β)/IL-1F2 Quantikine ELISA Kit (CAT#: MLB00C) and Mouse IL-6 Quantikine ELISA Kit (CAT#: M6000B) were purchased from R&D Systems, Inc. (Minneapolis, MN, Canada). MojoSort™ Human Pan Monocyte Isolation Kit (CAT#: 480060) was purchased from BioLegend (USA). Human Macrophage Colony-Stimulating Factor (M-CSF) Recombinant Protein (CAT#: RP-8643), Pierce™ BCA Protein Assay Kit (CAT#: 23227) and Amplex Red Hydrogen Peroxide⁄Peroxidase Assay Kit (CAT#: A22188) were purchased from Invitrogen (Carlsbad, CA, USA). Lactate Dehydrogenase (LDH) Assay Kit (CAT#: A020-2), Myeloperoxidase (MPO) Assay Kit (CAT#: A044-1-1), ROS Assay Kit (CAT#: E004-1-1), Malondialdehyde (MDA) Assay Kit (CAT#: A003-1-1), 4-Hydroxynonenal ELISA Kit (CAT#: H268), Protein Carbonyl Assay Kit (CAT#: A087-1-1), Total Anti-oxidant Capacity Assay Kit (CAT#: A015-2-1) and Superoxide Dismutase (SOD) Assay Kit (CAT#: A001-3-2) were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). TransAM® NF-κB p65 Kit was purchased from Active Motif (Carlsbad, CA, USA).
Protein carbonyl assay kit
Manufactured by Nanjing Jiancheng
Sourced in China
The Protein Carbonyl Assay Kit is a laboratory tool used to quantify the level of protein carbonylation in biological samples. The kit provides reagents and protocols to measure the oxidative modification of proteins, which is a key indicator of oxidative stress.
Automatically generated - may contain errors
Lab products found in correlation
Malondialdehyde mda assay kit, by Nanjing Jiancheng (2 mentions)
Superoxide dismutase sod assay kit, by Nanjing Jiancheng (2 mentions)
Mouse il 6 quantikine elisa kit, by R&D Systems (1 mentions)
Invivomab anti mouse ly6g, by BioXCell (1 mentions)
Mojosort human pan monocyte isolation kit, by BioLegend (1 mentions)
Lucigenin, by Merck Group (1 mentions)
Transam nf κb p65 kit, by Active Motif (1 mentions)
Lps e coli o111 b4, by Merck Group (1 mentions)
Amplex red hydrogen peroxide peroxidase assay kit, by Thermo Fisher Scientific (1 mentions)
Ros assay kit, by Nanjing Jiancheng (1 mentions)
Lactate dehydrogenase ldh assay kit, by Nanjing Jiancheng (1 mentions)
Total antioxidant capacity assay kit, by Nanjing Jiancheng (1 mentions)
Myeloperoxidase mpo assay kit, by Nanjing Jiancheng (1 mentions)
Reduced glutathione gsh assay kit, by Nanjing Jiancheng (1 mentions)
Folin ciocalteu phenol reagent, by Merck Group (1 mentions)
Sulfuric acid h2so4, by Sinopharm (1 mentions)
D galactose, by Merck Group (1 mentions)
Vitamin e, by Merck Group (1 mentions)
1 1 diphenyl 2 picrylhydrazyl, by Merck Group (1 mentions)
2 2 azino bis 3 ethylbenzothiazoline 6 sulfonic acid, by Merck Group (1 mentions)
5 protocols using protein carbonyl assay kit
1
Inflammatory Pathway Activation Assays
2
Oxidative Stress Biomarkers in Cells
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After treatment with 200 µg/mL BSA or MB with or without 10 µM 4′-methoxyresveratrol for 24 h, lysates from the cell culture were prepared using a homogenizer (ULTRA-TURRAX, IKA, Staufen, Germany). The concentration of total protein was measured by a BCA protein assay kit. Protein carbonyl expressed as nmol per mg total protein measured by a Protein Carbonyl Assay Kit according to the manufacturer’s protocol (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). The content of advanced oxidation protein products (AOPP) was determined as described by our previous study [21 (link)]. In brief, the standard curve was draw by 0, 10, 20, 40, 80, and 100 µmol/L chloramine-T and 300 µL of protein lysates were used for sample detection. Seventy-five microliters of 1.16 M KI and 150 µL of acetic acid were added into the tubes which served as reactants with chloramine-T or samples. After that, their absorbance was measured by microplate reader at 340 nm immediately. The AOPP quantity was calculated with respect to chloramine-T and results showed in nmol of chloramine-T equivalent per mg total protein.
3
Phytochemical Analysis and Antioxidant Assays
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Sodium carbonate (Na2CO3), glacial acetic acid (CH3COOH), edible ethanol, absolute ethanol, potassium acetate (C2H3KO2), aluminum trichloride (AlCl3), potassium chloride (KCl), sodium acetate (CH3COONa), methanol, sulfuric acid (H2SO4), and potassium persulfate (K2S2O8) originated from Sinopharm Chemical Reagent Co. (Shanghai, China). Folin–Ciocalteu phenol reagent, 1,1-diphenyl-2-picrylhydrazyl (DPPH), ascorbic acid, 2,2’-Azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), vitamin E (VE), and D-galactose were offered by Sigma-Aldrich Chemicals (St. Louis, MO, USA). PUSH BIO-TECHNOLOGY (Chengdu, China) provided: gallic acid, rutin, epicatechin, vanillin, formic acid, (+)-catechin, (-)-epigallocatechin, 3,4-dihydroxybenzoic acid, quercetin, epigallocatechin gallate, procyanidin A2, procyanidin B2, procyanidin A1, procyanidin B1, and luteolin. A malondialdehyde (MDA) assay kit, a superoxide dismutase (SOD) assay kit, a reduced glutathione (GSH) assay kit, and a protein carbonyl assay kit originated from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Pentobarbitol sodium was provided by HeFei BoMei Biotechnology Co. Ltd. (Hefei, China).
4
Assessing Muscle Protein Oxidation
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Carbonyl content was measured by the protein carbonyl assay kit (Jiancheng Bioengineering Institute, Nanjing, China). TCA soluble peptides were determined by the method of Jiang et al. (2021) .
The MFI was measured according to the method of Jiang et al. (2021) with slight modifications. One gram of minced muscle was homogenized by adding 15 mL pre-cooled MFI buffer (100 mM KCl, 11.2 mM K2HPO4, 8.8 mM KH2PO4, 1 mM EGTA, 1 mM MgCl2), passing a 200mesh filter, centrifuged at 1000×g for 15 min at 2 °C and the supernatant was discarded. Add again 15 mL MFI buffer, and repeat the above operation. Finally, the precipitate was adjusted to 0.5 mg/mL with MFI buffer, measuring the absorbance at 562 nm with an enzyme labeling measuring instrument (Sunrise-basic Tacan, TECAN, Swiss). The average absorbance times 200 was regarded as the MFI value.
The MFI was measured according to the method of Jiang et al. (2021) with slight modifications. One gram of minced muscle was homogenized by adding 15 mL pre-cooled MFI buffer (100 mM KCl, 11.2 mM K2HPO4, 8.8 mM KH2PO4, 1 mM EGTA, 1 mM MgCl2), passing a 200mesh filter, centrifuged at 1000×g for 15 min at 2 °C and the supernatant was discarded. Add again 15 mL MFI buffer, and repeat the above operation. Finally, the precipitate was adjusted to 0.5 mg/mL with MFI buffer, measuring the absorbance at 562 nm with an enzyme labeling measuring instrument (Sunrise-basic Tacan, TECAN, Swiss). The average absorbance times 200 was regarded as the MFI value.
5
Magnetic Iron Oxide Nanoparticles Synthesis
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Iron (III) chloride hexahydrate (FeCl3·6H2O), sodium oleate, oleic acid, 1-octadecene, poly (allylamine hydrochloride) (PAH; Mw = 15,000 Da), phosphate buffered saline (PBS), tetramethylammonium hydroxide, N-(3-(dimethylamino)propyl-N′-ethylcarbodiimide) hydrochloride (EDC), N-hydroxysulfosuccinimide sodium salt (NHS), 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2 H-tetrazolium bromide (MTT) assay kit and 2ʹ,7ʹ-dichlorohydrofluorescein diacetate (DCFH; ≥ 94%), glucose oxidase (GOx) were purchased from Sigma-Aldrich. 3,3′,5,5′-tetramethylbenzidine (TMB), 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), o-phenylenediamine (OPD; 98%), methylene blue (MB), terephthalic acid (TA) and hydrogen peroxide (H2O2) assay kit were obtained from Beyotime Biotechnology. Bicinchoninic acid (BCA) protein assay kit, malondialdehyde (MDA) and protein carbonyl assay kit were obtained from Nanjing Jiancheng Institute of Biological Engineering. Live/dead bacteria viability kit was purchased from Thermo Fisher Scientific. All other chemicals were obtained from Adamas-beta and used without further purification. Deionized (DI) water (Millipore Milli-Q grade, 18.2 MΩ) was used in all the experiments.
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