02 Oxidative Stress

MDA Assay Kit

MDA(malondialdehyde) Assay Kit

Capable of measuring MDA in cells and tissues
Simple procedure

Product code

M496　 MDA Assay Kit

Unit size	Price	Item Code
100 tests		M496-10

<Approximate number of uses> One plate of 96-well plate

Component

100 tests	・Lysis Buffer ・Dilution Buffer ・Standard ・Substrate ・Antioxidant	6.5 ml×1 10 ml×1 200 µl×1 58 mg×1 200 µl×1

C of A

SDS download

Description

　Malondialdehyde (MDA) is one of the end products of lipid peroxides, the MDA concentration in cell or tissue samples can indicate lipid peroxidation which is important in the fields of oxidative stress and ferroptosis. MDA is also called reactive aldehyde and reacts with amino and thiol groups to cause protein denaturation and DNA damage, so it is also measured in various disease studies such as cancer and diabetes.
　Dojindo's MDA Assay Kit uses the TBARS method to detect MDA by measuring the fluorescence or absorbance of the adduct of MDA and thiobarbituric acid that develops color according to the amount of MDA.

Manual

Manual

Technical info

The fluorometric method can measure both cell samples and tissue samples, the colorimetric method can only measure tissue samples. Please choose the proper method according to the expected concentration of samples.

	Fluorometric	Colorimetric	Amount of Sample	Detection range
Cell	OK	×	1-3×10⁷ cells	1-10 µmol/l
Tissue	OK	OK	Fluorometric: 10-30 mg Colorimetric: 20-50 mg	Fluorometric: 1-10 µmol/l Colorimetric: 1-50 µmol/l

Fluorometric

Colorimetric

Amount of Sample

Detection range

Cell

1-3×10⁷ cells

1-10 µmol/l

Tissue

Fluorometric: 10-30 mg

Colorimetric: 20-50 mg

Fluorometric: 1-10 µmol/l

Colorimetric: 1-50 µmol/l

Simple Procedures

Other company's TBA method MDA assay kit requires a substrate weighing procedure. however, Dojindo's Kit doesn't need a weighing procedure, thus greatly reducing the measuring time and the variation of assay results due to weighing errors.

Applicaiton Data: MDA level in Ferroptosis induced Cells

　Using Dojindo's MDA Assay Kit measuring the changes of MDA level in erastin (an inhibitor of xCT, Cystine/Glutamate transporter) treated HepG2 cells, the result shows that the GSSH/GSH ratio is decreased and the MDA, intracellular Fe²⁺, ROS, Lipid peroxide level is increased.

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References

Open References

No.	Sample	Reference (Link)
1	Tissue (Mouse Hippocampus)	K. Igarashi, H. Iwai, K. Tanaka, Y. Kuwahara, J. Kitanaka, N. Kitanaka, A. Kurimasa, K. Tomita, T. Sato, "Neuroprotective effect of oxytocin on cognitive dysfunction, DNA damage, and intracellular chloride disturbance in young mice after cranial irradiation", 2022, Biochem. Biophys. Res. Commun., doi:10.1016/j.bbrc.2022.04.099.
2	Cell (HeLa)	J. Zhu, X. Wang, Y. Su, J. Shao, X. Song, W. Wang, L. Zhong, L. Gan, Y. Zhao, X. Dong, "Multifunctional nanolocks with GSH as the key for synergistic ferroptosis and anti-chemotherapeutic resistance", 2022, doi:10.1016/j.biomaterials.2022.121704.
3	Cell (RAW 264.7)	B. Yang, G. Joe, W. Li, Y. Shimizu, H. Saeki, "Comparison of Maillard-Type Glycated Collagen with Alginate Oligosaccharide and Glucose: Its Characterization, Antioxidant Activity, and Cytoprotective Activity on H2O2-Induced Cell Oxidative Damage", 2022, doi:10.3390/foods11152374.
4	Cell (HepG2)	L. Dong, Z. Jiang, L. Yang, F. Hu, W. Zheng, P. Xue, S. Jiang, M. E. Andersen, G. He, M. J. C. Crabbe, W. Qu, "The genotoxic potential of mixed nitrosamines in drinking water involves oxidative stress and Nrf2 activation", 2022, doi:10.1016/j.jhazmat.2021.128010.
5	Cell (OMM-1)	C. Hou, L. Xiao, X. Ren, L. Cheng, B. Guo, M. Zhang, N. Yan., "EZH2-mediated H3K27me3 is a predictive biomarker and therapeutic target in uveal melanoma", 2022, Front. Genet., doi:10.3389/fgene.2022.1013475.
6	Cell (Human gingival fibroblasts)	L. Xing, W. Dong, Y. Chen, W. Dai, X. Xiao, Z. Liu, X. Zhang, D. Bai, H. Xu, "Fibroblast ferroptosis is involved in periodontitis-induced tissue damage and bone loss", 2022, Int. Immunopharmacol., doi:10.1016/j.intimp.2022.109607.
7	Cell (Huh7)	Y. Li, W. Yang, Y. Zheng, W. Dai, J. Ji, L. Wu, Z. Cheng, J. Zhang, J. Li, X. Xu, J. Wu, M. Yang, J. Feng and C. Guo, "Targeting fatty acid synthase modulates sensitivity of hepatocellular carcinoma to sorafenib via ferroptosis", J. Exp. Clin. Cancer Res., 2023, doi:10.1186/s13046-022-02567-z.
8	Tissue (Rat liver)	H. Liu, F. Yokoyaa, S. Ishizuka, "Metabolic alterations of the gut–liver axis induced by cholic acid contribute to hepatic steatosis in rats", Biochim. Biophys. Acta, Mol. Cell Biol. Lipids, 2023, doi:10.1016/j.bbalip.2023.159319.
9	Cell (4T1)	J. Zhao, Y. Chen, T. X, S. Han, C. Li, Y. He, Y. He, G. Zhao, G. Zhao, T. Wang, L. Wang, T. Cheng, C. Wang and J. Wang, "Clustered Cobalt Nanodots Initiate Ferroptosis by Upregulating Heme Oxygenase 1 for Radiotherapy Sensitization", Small, 2023, doi:10.1002/smll.202206415.
10	Tissue (Mouse pancreas)	L. Liu, Y. Xie, G. Li, T. Zhang, Y. Sui, Z. Zhao, Y. Zhang, W. Yang, X. Geng, D. Xue, H. Chen, Y. Wang, T. Lu, L. Shang, Z. Li, L. Li, B. Sun, "Gut microbiota-derived nicotinamide mononucleotide alleviates acute pancreatitis by activating pancreatic SIRT3 signalling", Br. J. Pharmacol., 2023, doi:10.1111/bph.15980.
11	Cell (Breast Cancer Cell)	Z. ZHu, H. Shen, J. Xu, Z. Fang, G. Wo, Y. Ma, K. Yang, Y. Wang, Q. Yu, J. Tang., "GATA3 mediates doxorubicin resistance by inhibiting CYB5R2-catalyzed iron reduction in breast cancer cells", 2023, doi:10.1016/j.drup.2023.100974.

Q & A

Q What is the principle of this kit?: A

This kit uses 2-thiobarbituric acid(TBS) as a substrate. By measuring the absorbance or fluorescence intensity of the TBA adduct formed by the reaction of MDA and TBA, and comparing with the standard curve, the MDA concentration in the sample can be simply measured.

Q How many samples can be tested by this kit?: A

When testing the sample in triplicates, 24 samples can be tested by this kit.

Q What is the storage condition of the prepared solution?: A

Substrate stock solution is stable for 2 months under -20℃.
Antioxidant PBS solution and Working solution is stable for 24 hours.

Q Do I need to normalize the result by cell numbers?: A

If there is no change in cell number after drug stimulation, the normalization by cell number is not necessary.
If the number of cells changes, the normalized result by protein quantification is more accurate, in such case, the BCA method is recommended for protein quantification.

Handling and storage condition

Handling and storage condition
1. Hazardous material Class 4, Petroleums No. 2, Hazardous material Class III, Petroleums No. 3, Hazardous material Class III, 2. Health and Safety Law, Chemical Substances Control Law, 3. Prevent from fire. 4. Store in a deep freeze at -20℃.
PRTR Policy	Class 1 designated chemical substance
Danger / harmful symbol mark