Amplite® Colorimetric L-Lactate Dehydrogenase (LDH) Assay Kit
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Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Overview | ![]() ![]() |
Platform
Absorbance microplate reader
Absorbance | 575/605 nm |
Recommended plate | Clear bottom |
Components
Example protocol
AT A GLANCE
- Prepare L-Lactate Dehydrogenase standards or test samples (50 µL)
- Add L-Lactate Dehydrogenase working solution (50 µL)
- Incubate at room temperature for 30 min - 2 hours
- Monitor absorbance ratio increase at A575nm/A605nm
Thaw one vial of each kit component at room temperature before starting the experiment.
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles
Add 100 µL of H2O into the vial of NAD (Component C) to make 100X NAD stock solution.
Add 100 µL of H2O or 1x PBS buffer into the vial of L-Lactate Dehydrogenase (Component D) to make 100 U/mL L-LDH standard solution.
PREPARATION OF STANDARD SOLUTIONS
https://www.aatbio.com/tools/serial-dilution/13813
PREPARATION OF WORKING SOLUTION
- Add 10 mL of Assay Buffer (Component B) into the bottle of Enzyme Mix (Component A), and mix well.
Add 100 µL of 100X NAD stock solution into the bottle of Component A+B and mix well to make L-LDH working solution.
Note This L-LDH working solution is enough for two 96-well plates. It is unstable and should be used promptly within 2 hours. Avoid exposure to light.
Note Alternatively, one can make a 50X of L-LDH Enzyme Mix stock solution by adding 200 μL of H2O into the bottle of Enzyme Mix (Component A), and then prepare the L-LDH working solution by mix the stock solution with Assay Buffer (Component B) and 100X NAD stock solution proportionally.
SAMPLE EXPERIMENTAL PROTOCOL
Table 1. Layout of L-LDH standards and test samples in a white clear bottom 96-well microplate. SD= L-LDH Standards (SD1 - SD7, 0.4 to 300 mU/mL), BL=Blank Control, TS=Test Samples.
BL | BL | TS | TS |
SD1 | SD1 | ... | ... |
SD2 | SD2 | ... | ... |
SD3 | SD3 | ||
SD4 | SD4 | ||
SD5 | SD5 | ||
SD6 | SD6 | ||
SD7 | SD7 |
Table 2. Reagent composition for each well.
Well | Volume | Reagent |
SD1 - SD7 | 50 µL | Serial Dilutions (0.4 to 300 mU/mL) |
BL | 50 µL | Dilution Buffer |
TS | 50 µL | test sample |
- Prepare L-LDH standards (SD), blank controls (BL), and test samples (TS) according to the layout provided in Tables 1 and 2. For a 384-well plate, use 25 µL of reagent per well instead of 50 µL.
- Add 50 µL of L-LDH working solution to each well of L-LDH standard, blank control, and test samples to make the total L-LDH assay volume of 100 µL/well. For a 384-well plate, add 25 µL of L-LDH working solution into each well instead, for a total volume of 50 µL/well.
- Incubate the reaction at room temperature for 30 minutes to 2 hours, protected from light.
- Monitor the absorbance ratio increase with an absorbance plate reader at A575nm/A605nm.
Images
Citations
Authors: Brown, Aric
Journal: (2023)
Authors: Xia, Peng and Zhang, Hao and Lu, Haofeng and Xu, Kequan and Jiang, Xiang and Jiang, Yuke and Gongye, Xiangdong and Chen, Zhang and Liu, Jie and Chen, Xi and others,
Journal: Cancer Communications (2023)
Authors: Deka, Ranjit K and Liu, Wei Z and Norgard, Michael V and Brautigam, Chad A
Journal: Mbio (2020): e02249--20
Authors: Han, Dong and Wang, Yong-Jun and Chen, Jiangwei and Zhang, Jibin and Yu, Peng and Zhang, Ran and Li, Shuang and Tao, Bo and Wang, Yabin and Qiu, Ya and others, undefined
Journal: Journal of pineal research (2019): e12571
Authors: Song, Jin H and Singh, Neha and Luevano, Libia A and Padi, Sathish KR and Okumura, Koichi and Olive, Virginie and Black, Stephen M and Warfel, Noel A and Goodrich, David W and Kraft, Andrew S
Journal: Molecular cancer therapeutics (2018): 2710--2721
Authors: Wang, Yujue and Cao, Weifeng and Luo, Jianquan and Qi, Benkun and Wan, Yinhua
Journal: Bioresource Technology (2018)
Authors: Chen, Liping and Mai, Weiqian and Chen, Minfeng and Hu, Jianyang and Zhuo, Zhenjian and Lei, Xueping and Deng, Lijuan and Liu, Junshan and Yao, Nan and Huang, Maohua and others, undefined
Journal: Pharmacological Research (2017)
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Journal: Journal of Medicinal Chemistry (2017)
Authors: Zhang, Xiaofang and Wang, Lei and Zhang, Xiaodong and Ren, Lijun and Shi, Wenjing and Tian, Yijun and Zhu, Jiangbo and Zhang, Tianbao
Journal: Food and Chemical Toxicology (2017)
Authors: Sünwoldt, Juliane and Bosche, Bert and Meisel, Andreas and Mergenthaler, Philipp
Journal: Frontiers in Molecular Neuroscience (2017): 305
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Application notes
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