LDH Assay Kit (Cytotoxicity) (ab65393)
Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 1 hr
- Sample type: Adherent cells, Suspension cells
製品の概要
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製品名
LDH Assay Kit (Cytotoxicity)
Lactate Dehydrogenase キット 製品一覧 -
検出方法
Colorimetric -
サンプルの種類
Adherent cells, Suspension cells -
アッセイタイプ
Enzyme activity (quantitative) -
全工程の試験時間
1h 00m -
製品の概要
LDH Assay Kit (Cytotoxicity) ab65393 uses WST for the fast and sensitive detection of LDH released from damaged cells.
The LDH assay, also known as LDH release assay, is a cell death / cytotoxicity assay used to assess the level of plasma membrane damage in a cell population. Lactate dehydrogenase (LDH) is a stable enzyme, present in all cell types, which is rapidly released into the cell culture medium upon damage of the plasma membrane. LDH is the most widely used marker used to run a cytotoxicity assay.
How the assay works
The LDH assay protocol is based on an enzymatic coupling reaction: LDH released from the cell oxidizes lactate to generate NADH, which then reacts with WST to generate a yellow color. The intensity of the generated color correlates directly with the number of lyzed cells.
Only 10μl of culture medium is required for the assay, and thus background from serum and culture medium is significantly reduced. Cells can be cultured in regular 10% serum containing medium; no reducing serum or special medium is required for the assay.
This LDH assay kit uses a chemical method to couple the NADH reaction with WST, which is more tolerant of variability in the sample than the diaphorase-based coupling method used in kits available from other vendors. The chemical method means that the active ingredients of the kit, excluding the LDH positive control, do not contain enzymes, meaning they are more stable for practical use in the lab in repeated experiments.
In addition, the WST used in the kit is highly stable, soluble in solution and unlikely to precipitate in solution, unlike the INT used in kits available from other vendors. This means that the reaction can be read multiple times over a long time-course. The reaction can also be stopped at any time point.
LDH assay protocol summary
- Transfer 10μl culture medium into new plate
- Add LDH reaction mix and incubate for 30 min at room temp
- Analyze with microplate reader
LDH activity can be easily quantified by spectrophotometer or plate reader at OD450nm.
Related LDH assay products
If you would like to use a fluorometric assay, please refer to LDH-Cytotoxicity Assay Kit (Fluorometric) ab197004.
This kit is more sensitive than the colorimetric LDH Cytoxicity Assay Kit ab65391.
To measure LDH activity in sample types such as serum, plasma, and cell lysates, we recommend LDH assay kit ab102526.
Other notes
This product is manufactured by BioVision, an Abcam company and was previously called K313 LDH-Cytotoxicity Colorimetric Assay Kit II. K313-500 is the same size as the 500 test size of ab65393. -
特記事項
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the SDS download section.
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試験プラットフォーム
Microplate reader
製品の特性
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保存方法
Store at -20°C. Please refer to protocols. -
内容 500 tests 10000 tests Lysis Buffer II 1 x 5ml 20 x 5ml LDH Positive Control 1 vial 20 vials LDH Assay Buffer 1 x 50ml 20 x 50ml Stop Solution IV 1 x 5ml 20 x 5ml Substrate Mix 1 vial 20 vials -
研究分野
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パスウェイ
Fermentation; pyruvate fermentation to lactate; (S)-lactate from pyruvate: step 1/1. -
関連疾患
Defects in LDHA are the cause of glycogen storage disease type 11 (GSD11) [MIM:612933]. A metabolic disorder that results in exertional myoglobinuria, pain, cramps and easy fatigue. -
配列類似性
Belongs to the LDH/MDH superfamily. LDH family. -
翻訳後修飾
ISGylated. -
細胞内局在
Cytoplasm. - Information by UniProt
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別名
- Cell proliferation-inducing gene 19 protein
- GSD11
- L lactate dehydrogenase B chain
see all
関連製品
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Assay kits
画像
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LDH Cytotoxicity Assay using ab65393Image from Bukong TN et al., PLoS Pathog 10(10), Fig S6. Doi: 10.1371/journal.ppat.1004424. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Bafilomycin A1 (BafA1) toxicity was assessed in Human hepatic (Huh7.5) cells after 24 hours at different concentrations (12.5nm, 25nm, 50nm and 100nm) were administered to cells using LDH cytotoxicity assay kit (ab65393). Cytotoxicity was measured by subtracting LDH content in remaining viable cells from total LDH in untreated controls.
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LDH Cytotoxicity Assay useing ab65393Jurkat T cells were cultured in 96-well plate in 100 μl of culture medium. LDH Assay was performed using 10μl of culture medium using the WST probe. Low control (white bar); High control (black bar).
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LDH Cytotoxicity Assay using ab65393Comparison of WST-1 and INT based LDH assays. 3T3 cells were cultured in a 96-well plate in 100 µl of culture medium. The LDH assay was performed using 10 µl of culture medium using WST-1 (Brown bar) and INT (Green bar) methods. The WST-1 based LDH assay is more stable and sensitive than the INT based method.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (152)
ab65393 は 152 報の論文で使用されています。
- Fertan E et al. Cerebral organoids with chromosome 21 trisomy secrete Alzheimer's disease-related soluble aggregates detectable by single-molecule-fluorescence and super-resolution microscopy. Mol Psychiatry 29:369-386 (2024). PubMed: 38102482
- Li T et al. Deficiency of inflammation-sensing protein neuropilin-2 in myeloid-derived macrophages exacerbates colitis via NF-κB activation. J Pathol 262:175-188 (2024). PubMed: 37946610
- Shtuhin-Rahav R et al. Enteropathogenic E. coli infection co-elicits lysosomal exocytosis and lytic host cell death. mBio 14:e0197923 (2023). PubMed: 38038448
- Haritan N et al. Topology and function of translocated EspZ. mBio 14:e0075223 (2023). PubMed: 37341483
- Tomlinson KL et al. Staphylococcus aureus stimulates neutrophil itaconate production that suppresses the oxidative burst. Cell Rep 42:112064 (2023). PubMed: 36724077