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4 hne assay kit

Manufactured by Abcam
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The 4-HNE assay kit is a quantitative colorimetric assay designed to measure the levels of 4-hydroxynonenal (4-HNE), a lipid peroxidation marker, in biological samples. The kit provides a simple and reliable method to detect and quantify 4-HNE in a variety of sample types.

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Lab products found in correlation

Mda assay kit, by Nanjing Jiancheng (3 mentions) Mda assay kit, by Abcam (2 mentions) Pierce bca protein kit, by Thermo Fisher Scientific (2 mentions) Epoch microplate reader, by Agilent Technologies (2 mentions) Sod assay kit, by Nanjing Jiancheng (1 mentions) A003 1 2, by Nanjing Jiancheng (1 mentions) Quantichrom gsh assay kit, by BioAssay Systems (1 mentions) Mda assay kit, by Merck Group (1 mentions) Spectramax paradigm multi mode microplate reader, by Molecular Devices (1 mentions) Microplate reader, by Agilent Technologies (1 mentions) Lpo mda assay kit, by Beyotime (1 mentions) Nadph oxidase assay kit, by Nanjing Jiancheng (1 mentions) Gsh and gssg assay kit, by Beyotime (1 mentions) Protease inhibitor, by Thermo Fisher Scientific (1 mentions) Phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions) 3 nt elisa kit, by Abcam (1 mentions)

14 protocols using 4 hne assay kit

1

Oxidative Stress Biomarkers Quantification

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Levels of malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) were assessed using the MDA assay kit (Jiancheng Bioengineering Institute, Nanjing, China) and the 4-HNE assay kit (Abcam, Cambridge, UK) according to the manufacturer’s protocols. Superoxide dismutase (SOD) activity was determined using the SOD assay kit (Jiancheng Bioengineering Institute, Nanjing, China) following the manufacturer’s protocol.
Yuan Y., Zhai Y., Chen J., Xu X, & Wang H. (2021). Kaempferol Ameliorates Oxygen-Glucose Deprivation/Reoxygenation-Induced Neuronal Ferroptosis by Activating Nrf2/SLC7A11/GPX4 Axis. Biomolecules, 11(7), 923.
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2

Detecting Oxidative Stress in Spermatozoa

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Spermatozoa were separated from seminal plasma by centrifugation for the detection of ROS by 2,7-dichlorofluorescin diacetate (DCFH-DA) as described previously [48 (link)]. Cells were incubated with DCFH-DA (10 μmol/L) at 37 °C for 60 min, and ROS production was determined by a fluorescence microplate reader. 4-Hydroxynonenal (4-HNE)-protein adducts were detected using a Lipid Peroxidation (4-HNE) Assay Kit (Abcam, ab238538). This competitive ELISA kit allowed detection of the 4-HNE adduct in testis samples determined by comparing its absorbance with that of a known 4-HNE-BSA standard curve.
Mu Y., Dai H.G., Luo L.B, & Yang J. (2021). Irisin alleviates obesity-related spermatogenesis dysfunction via the regulation of the AMPKα signalling pathway. Reproductive Biology and Endocrinology : RB&E, 19, 135.
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3

Oxidative Stress Biomarker Assessment

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The content of malondialdehyde (MDA) was detected by its commercially available kit (A003-1-2) from the Jiancheng Bioengineering Institute (Nanjing, China). The expression level of 4-hydroxy-nonenal (4-HNE) in the cell lysate or tissue lysate was detected using the Lipid Peroxidation (4-HNE) Assay Kit (ab238238, Abcam). The ROS content was detected using its commercial kit (JL13783) from Shanghai Jianglai Biological Technology Co., Ltd. (Shanghai, China).
Wu W., Zhang G., Qiu L., Liu X., Zhou S, & Wu J. (2022). Contribution of Adiponectin/Carnitine Palmityl Transferase 1A-Mediated Fatty Acid Metabolism during the Development of Idiopathic Pulmonary Fibrosis. Oxidative Medicine and Cellular Longevity, 2022, 5265616.
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4

Quantification of Oxidative Stress Markers

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The relative MDA concentration in cell or tumor lysates was assessed using a Lipid Peroxidation (MDA) Assay Kit (ab118970, Abcam), according to the manufacturer’s instructions. This assay measures MDA reaction with thiobarbituric acid (TBA) that generate a MDA-TBA adduct in a sample. The MDA-TBA adduct can be quantified colorimetrically (OD = 532 nm). Lipid Peroxidation (4-HNE) Assay Kit (Abcam, ab238538) were used to evaluate the concentration of 4-HNE according to the manufacturer’s protocol.
Li P., Lin Q., Sun S., Yang N., Xia Y., Cao S., Zhang W., Li Q., Guo H., Zhu M., Wang Y., Zheng Z, & Li S. (2022). Inhibition of cannabinoid receptor type 1 sensitizes triple-negative breast cancer cells to ferroptosis via regulating fatty acid metabolism. Cell Death & Disease, 13(9), 808.
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5

Ferroptosis Markers Quantification

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The level of MDA, 4-HNE and GSH were measured to evaluate ferroptosis in different groups. Herein, the concentration of MDA, 4-HNE and GSH in tissue lysates were measured with the MDA Assay Kit (MAK085, Sigma-Aldrich), 4-HNE Assay Kit (ab238538, Abcam), and QuantiChrom™ GSH Assay Kit (DIGT250, Bioassay Systems) according to the manufacturer’s instructions respectively.
Liu P., Yuan J., Feng Y., Chen X., Wang G, & Zhao L. (2021). Ferroptosis contributes to isoflurane-induced neurotoxicity and learning and memory impairment. Cell Death Discovery, 7, 72.
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6

Quantifying Lipid Peroxidation in Cells and Lung

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The levels of 4HNE in cell culture supernatant and lung homogenate were measured using a lipid peroxidation (4HNE) assay kit (Abcam, Cambridge, MA).
Marshall R.J., Armart P., Hulme K.D., Chew K.Y., Brown A.C., Hansbro P.M., Bloxham C.J., Flint M., Ronacher K., Bielefeldt-Ohmann H., Gallo L.A, & Short K.R. (2020). Glycemic Variability in Diabetes Increases the Severity of Influenza. mBio, 11(2), e02841-19.
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7

Quantifying Oxidative Stress in Brown Adipose Tissue

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10 mg of brown adipose tissue (BAT) samples were homogenized with a dounce homogenizer and 200 μL RIPA buffer. Samples were centrifuged at 4 °C and pelleted down. Supernatants were centrifuged again at 4 °C, lysates were frozen at −80 °C. Lysates were diluted 2× and run with a 4-hydroxynonenal (4-HNE) assay kit (ab238538, Abcam, Cambridge, MA) on an Epoch microplate reader (BioTek Inc., Winooski, VT) at an absorbance of 450 nm in duplicates and results were plotted along a standard curve. Lysate protein concentration was calculated with a Pierce BCA Protein kit (ThermoScientific, Rockford, Il). Lysates were diluted by 4× and 2% SDS was added to samples to limit interference by lipids (Kessler and Fanestil 1986 (link)). Samples were measured on an Epoch microplate reader (BioTek Inc., Winooski, VT) at an absorbance of 562 nm in duplicates and results were plotted along a standard curve. We selected BAT to measure 4-HNE due to its high metabolic activity in hibernation. Recent data may indicate metabolically active tissues such as BAT might have higher levels of molecular damage over hibernation in AGS (Wilbur et al. 2019 (link)) and our lab has shown BAT is the only tissue to show oxidative stress after arousal (Orr et al. 2009 (link)).
Mikes M., Rice S.A., Bibus D., Kitaysky A, & Drew K.L. (2022). Translating PUFA omega 6:3 ratios from wild to captive hibernators (Urocitellus parryii) enhances sex-dependent mass-gain without increasing physiological stress indicators. Journal of Comparative Physiology. B, Biochemical, Systemic, and Environmental Physiology, 192(3-4), 529-540.
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8

Lipid Peroxidation Quantification

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The concentration of MDA and 4-HNE was detected using lipid peroxidation (MDA) Assay kit (abcam, USA) and lipid peroxidation (4-HNE) Assay kit (abcam, USA) according to the manufacturer's instructions. The OD value or fluorescence signal was determined by SpectraMax Paradigm Multi-Mode Microplate Reader (MOLECULAR DEVICES, USA).
Wang L., Huang B., Zeng Y., Yang J., Li Z., Ng J.P., Xu X., Su L., Yun X., Qu L., Chen R., Luo W., Wang Y., Chen C., Yang L., Qu Y., Zhang W., Chan J.T., Wang X., Law B.Y., Mok S.W., Chung S.K, & Wong V.K. (2023). N-Acetylcysteine overcomes epalrestat-mediated increase of toxic 4-hydroxy-2-nonenal and potentiates the anti-arthritic effect of epalrestat in AIA model. International Journal of Biological Sciences, 19(13), 4082-4102.
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9

Quantifying Lipid Peroxidation Markers

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MDA levels in cells were measured using an LPO MDA assay kit (Beyotime Biotechnology). Cellular proteins were extracted and added to MDA detection working solution and then heated in boiling water for 15 min. The absorbance was measured at 532 nm using a microplate reader to calculate the MDA levels. 4-HNE levels were determined using a 4-HNE assay kit (Abcam Inc., Cambridge, MA, USA) according to the manufacturer’s instructions, and the OD values of the cells were determined using a microplate reader (Bio-Tek Instruments, Winooski, VT, USA) at 450 nm. Then, a standard curve was drawn based on the OD values of the standard, and the concentration of the samples was calculated.
Du Y.X., Zhao Y.T., Sun Y.X, & Xu A.H. (2023). Acid sphingomyelinase mediates ferroptosis induced by high glucose via autophagic degradation of GPX4 in type 2 diabetic osteoporosis. Molecular Medicine, 29, 125.
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10

Myocardial Lipid Peroxidation Analysis

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Lipid peroxidation levels in the myocardial tissue and PAMCs were measured by detecting malondialdehyde (MDA) and 4-hydroxynonenalal (4-HNE) contents using an MDA Assay Kit (A003, Jiancheng) and a 4-HNE Assay Kit (ab233471, Abcam), respectively, according to the manufacturers’ instructions [8 (link)].
Wang L., Qiao Y., Yu J., Wang Q., Wu X., Cao Q., Zhang Z., Feng Z, & He H. (2024). Endurance exercise preconditioning alleviates ferroptosis induced by doxorubicin-induced cardiotoxicity through mitochondrial superoxide-dependent AMPKα2 activation. Redox Biology, 70, 103079.
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