Astragaloside IV (> 98%) was purchased from Nanjing Spring & Autumn Biological Engineering (Nanjing, China) and stored in the dark at −20°C and then dissolved in dimethyl sulfoxide (DMSO) (Solarbio, Beijing, China). It was also diluted with complete medium prior to experimentation. H2O2 and t-BHQ were obtained from Sigma-Aldrich (St. Louis, MO, USA). Penicillin and streptomycin, Fetal bovine serum (FBS), and Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 (DMEM/F-12) were obtained from Invitrogen-Gibco (Grand Island, NY, USA). Primary antibodies used targeted NFE2L2 (16396-1-AP, Proteintech, Chicago, IL, USA), NQO1 (11451-1-AP, Proteintech, Chicago, IL, USA), HMOX1 (ab13248, Abcam, Cambridge, UK), and β-actin (GB12001, Servicebio, Wuhan, China). GSH-Px, SOD, CAT, T-AOC, and MDA test kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).
Mda test kit
Manufactured by Nanjing Jiancheng
Sourced in China
The MDA test kit is a laboratory equipment used for the detection and quantification of MDA (Malondialdehyde), a biomarker commonly associated with oxidative stress and certain disease states. The kit provides a standardized method for analyzing MDA levels in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Microplate reader, by Thermo Fisher Scientific (4 mentions)
Gsh px, by Nanjing Jiancheng (2 mentions)
T aoc, by Nanjing Jiancheng (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Iron assay kit, by Nanjing Jiancheng (2 mentions)
Sod test kit, by Nanjing Jiancheng (2 mentions)
Penicillin and streptomycin, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium nutrient mixture f 12 dmem f 12, by Thermo Fisher Scientific (1 mentions)
Hmox1, by Abcam (1 mentions)
β actin, by Wuhan Servicebio Technology (1 mentions)
Nfe2l2 16396 1 ap, by Proteintech (1 mentions)
Nqo1 11451 1 ap, by Proteintech (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Solarbio (1 mentions)
Glutathione (gsh), by Nanjing Jiancheng (1 mentions)
Reverse transcription kit, by Takara Bio (1 mentions)
Taq pcr starmix, by GenStar (1 mentions)
Sodium dodecyl sulfate (sds), by Biosharp (1 mentions)
Tb green premix ex taq 2 2, by Takara Bio (1 mentions)
E z n a mollusc dna kit, by Omega Bio-Tek (1 mentions)
21 protocols using mda test kit
1
Astragaloside IV Antioxidant Pathway
2
Endogenous Antioxidant Status Determination
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Serum was isolated in the form of clear supernatant which was further utilized for determination of endogenous antioxidants status superoxide dismutase (SOD), catalase (CAT), malonaldehyde (MDA) and reduced glutathione (GSH). SOD, CAT, GSH and MDA test kits were purchased from Nanjing Jiancheng Bioengineering Institute (China). The assay for total SOD was based on its ability to inhibit the oxidation of oxymine by the xanthine–xanthine oxidase system.19 (link) The hydroxylamine nitrite produced by the oxidation of oxymine had an absorbance peak at 550 nm. One unit of SOD activity was defined as the amount that reduced the absorbance at 550 nm by 50%. CAT activity was assayed following the procedure which is based on the rate of decrease of H2O2 at 240 nm with detection limit of 0.002 U/mL.20 (link) Thiobarbituric acid reaction (TBAR) method was used to determine the MDA which can be measured at the wavelength of 532 nm by reacting with thiobarbituric acid (TBA) to form a stable chromophoric production.21 (link) Analysis of GSH was performed with the method using DTNB as a substrate at 412 nm with detection limit of 1.6 mmol/L.22 (link)
3
Comprehensive Biochemical and Molecular Analysis
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SDS was purchased from Biosharp Company of China, and the purity was 99%. Total protein quantitation kit, SOD, CAT, and MDA test kits were purchased from Nanjing Jiancheng Company of China for determination of the activities of SOD, CAT and MDA content. The E.Z.N.A.® Mollusc DNA kit was the product of Omega Bio-Tek Company for extraction of genomic DNA, and 2 × Taq PCR StarMix was purchased from GenStar Company. Trizol reagent was purchased from Thermo Fisher Technology Co., Ltd. for extraction of RNA. Reverse Transcription kit and TB Green premix Ex Taq II (2×) were purchased from TaKaRa Company. The sequences of 13 random primers and qPCR primers used in this study are shown in Tables S1 and S2 .
4
Germacrone-Induced Apoptosis Signaling
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In the present study, Germacrone (GM) (≥ 98%) was purchased from Nanjing Puyi Biological Technologies. 8oHdG and MDA test kits were obtained from the Nanjing Jiancheng Bioengineering Institute in Nanjing, China. Rabbit antibodies targeting proteins associated with apoptosis (Bcl-2, Bax, cleaved caspase-3, NF-κB, and COX-2) were purchased from Abcam (Cambridge, USA. Santa Cruz Biotechnology, Dallas, TX, USA). Rabbit tubulin and goat anti-rabbit IgG for peroxidase were acquired from Santa Cruz Biotechnology.
5
Biochemical Assays for Oxidative Stress
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The activities of T-AOC, SOD, GSH-Px, CAT, and MDA test kits were acquired from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). These tests were performed according to the manufacturer’s instructions.
6
Evaluating Tobacco Stress Responses
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Five-week-old potted tobacco seedlings were used for the salt and drought stress treatments. For inducing drought stress, we stopped watering tobacco plants for 10 days. The seedlings were irrigated with 200 mM NaCl solution to induce salt stress. Phenotypes before and after the stress treatments were photographed and preserved. Samples were collected to determine the physiological indicators. Chlorophyll and the relative electrolyte leakage (REL) were determined according to Dong et al. (2020) [30 (link)]. Malondialdehyde (MDA) levels were measured by the barbiturate method using an MDA test kit (Nanjing Jiancheng, Nanjing, China). The relative water loss was measured according to Gong et al. (2015) [28 (link)], as well as the DAB and NBT staining. The stress treatments were performed with at least three biological replicates with identical plant numbers in each replicate.
7
Serum Lipid and Oxidative Profiles
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The level of high-density lipoprotein (HDL), total cholesterol (TC), triglyceride (TG) and low-density lipoprotein (LDL) in serum were examined with an automatic biochemical analyzer (model 7150; Hitachi, Tokyo, Japan) according to the manufacturer’s instructions.
The concentration of superoxide dismutase (SOD) and MDA were measured using commercial total superoxide dismutase (T-SOD) test kit (A001-1-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China) and MDA test kit (A003-1-1, Nanjing Jiancheng Bioengineering Institute) according to the operating manual. The absorbance of wells was determined at 560 nm (SOD) and 532 nm (MDA) using a microplate reader (Thermo Fisher Scientific), respectively.
The concentration of superoxide dismutase (SOD) and MDA were measured using commercial total superoxide dismutase (T-SOD) test kit (A001-1-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China) and MDA test kit (A003-1-1, Nanjing Jiancheng Bioengineering Institute) according to the operating manual. The absorbance of wells was determined at 560 nm (SOD) and 532 nm (MDA) using a microplate reader (Thermo Fisher Scientific), respectively.
8
Serum Analysis After hUCMSC Transplantation
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Blood samples were obtained 3 h and 2 and 7 days after hUCMSC transplantation and centrifuged at 1000 ×g for 15 mins, and the serum was collected. Serum samples were tested for alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA), and total bilirubin (TBIL) using ALT/GPT microplate test kit, AST/GOT microplate test kit, MDA test kit, and TBIL test kit, respectively (Nanjing Jiancheng Bioengineering Institute).
9
Squid Mantle MDA Content Measurement
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The malondialdehyde (MDA) content in the squid mantle was measured using a MDA test kit (Nanjing Jiancheng Bio. Inst., NanJing, China). The squid mantle muscle was homogenized (1:9, w/v) in a normal saline solution, and the homogenates were centrifuged at 8,000 × g for 15 min at 4°C. The obtained supernatants were collected, and the MDA content was measured. The measurement was conducted according to the manufacturer’s instructions and expressed as nmol MDA per mg muscle.
10
Oxidative Stress Biomarkers in Lung
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GSH, MDA, 4HNE and Fe2+ in lung tissues and cells were respectively measured using a reduced GSH assay kit (A006-2-1), a MDA test kit (A003-1), a 4-HNE ELISA kit (H268) and an iron assay kit (A039-2-1) purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China), in accordance with the manufacturer’s instructions.
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