Determining Oxidative Damage by Lipid Peroxidation Assay in Rat Serum
利用脂质过氧化测定检测大鼠血清的氧化损伤
发布: 2019年06月20日第9卷第12期 DOI: 10.21769/BioProtoc.3263 浏览次数: 9685
评审: Davide BottaHua WangAnonymous reviewer(s)
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The authors used this protocol in:
Feb 2019
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Abstract
It has been well-established that malondialdehyde (MDA), which is generated during the process of lipid peroxidation, is a commonly known biomarker for oxidative stress. Therefore, the serum levels of MDA are detected by using the lipid peroxidation assay with commercially available kit to determine the induction of oxidative stress in rat models.
Keywords: Oxidative stress (氧化损伤)
Malondialdehyde (丙二醛)
Lipid peroxidation (脂质过氧化)
Thiobarbituric acid (硫代巴比妥酸)
MDA-TBA adduct (MDA-TBA加合物)
Serum (血清)
Background
As lipid peroxidation is the degradation of lipids that occurs as a result of oxidative damage and contributes to the pathology of many diseases, some end-products of the chain reaction of lipid peroxidation such as malondialdehyde (MDA), 4-Hydroxynonenal (4-HNE) and 8-iso-Prostaglandin F2alpha (8-isoprostane) in serum samples have been detected and quantified for the identification of oxidative damage (Marrocco et al., 2017). In the current lipid peroxidation assay protocol, the serum levels of MDA could be specifically and reliably quantified based on the condensation reaction between MDA and thiobarbituric acid (TBA) by modifying a commercially available kit (MAK085D; Sigma, St Louis, MO) without n-butanol precipitation step (Tang et al., 2019).
Materials and Reagents
- Pipette tips
- 96-well flat bottom microplate (Sigma-Aldrich, catalog number: CLS3610)
- Separate tube (Corning, catalog number: CLS430791)
- Double-distilled water (ddH2O)
- Ice
- 60 μl freshly collected rat serum
- TBA (Sigma-Aldrich, catalog number: MAK085D)
- Glacial acetic acid (Sigma-Aldrich, catalog number: A6283)
- 4.17 M MDA standard (Sigma-Aldrich, catalog number: MAK085E)
- 42 mM Sulfuric acid solution (Sigma-Aldrich, catalog number: 84736)
- Phosphotungstic acid solution (PTA) (Sigma-Aldrich, catalog number: MAK085B)
- Butylated hydroxytoluene (BHT) (Sigma-Aldrich, catalog number: MAK085C)
- MDA standard (see Recipes)
Equipment
- Pipettes (Bio-rad, P10, P200 and P1000)
- Vortexer (Ratek, catalog number: VM1)
- Centrifuge (Thermoline, model: K1015 Pro)
- Incubator (Robbins Scientific, catalog number: 2000)
- Microplate reader (Perkin Elmer, catalog number: 1420 Victor3)
Software
- Microsoft Excel 2010
Procedure
文章信息
版权信息
© 2019 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Tang, Q., Su, Y. and Xian, C. J. (2019). Determining Oxidative Damage by Lipid Peroxidation Assay in Rat Serum. Bio-protocol 9(12): e3263. DOI: 10.21769/BioProtoc.3263.
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分类
发育生物学 > 细胞信号传导 > 胁迫反应
细胞生物学 > 细胞新陈代谢 > 脂质
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