| Usage | 1. Preparation of cell culture medium samples to be tested:
(1) Take 1 mL of cell culture medium from a cell culture in the logarithmic growth phase.
(2) Centrifuge the cell culture medium at 200g for 5 minutes at room temperature. Transfer approximately 800 μL of the culture medium to a new microtube, taking care not to carry away any cell debris.
(3) Perform luciferase mycoplasma detection using the cell culture medium prepared above.
2. Luciferase mycoplasma detection:
(1) Take 50 μL of cell culture medium sample and add it to a 96-well white opaque cell plate.
(2) Equilibrate the luciferase mycoplasma detection reagent and substrate to room temperature (22℃-25℃) and gently shake to mix.
(3) Add 50 μL of luciferase mycoplasma detection reagent to the 50 μL cell culture medium sample.
(4) Vortex to mix for 10 seconds, incubate at room temperature for 5 minutes, and read the fluorescence value (reading 1).
(5) Add 10 μL of luciferase mycoplasma detection substrate.
(6) Vortex and mix for 10 seconds. Incubate at room temperature for 10 minutes and read the fluorescence value (reading 2).
3. Results
The ratio of reading 2 to reading 1 is used to determine whether there is mycoplasma contamination. | Ratio of reading 2 to reading 1 | Result interpretation and suggestions | | < 1 | No Mycoplasma contamination | | 1-1.2 | Possible Mycoplasma contamination. Isolate cells, culture for 24-48 hours and then retest for Mycoplasma contamination. | | Description | The MYCO-LTM Luciferase Mycoplasma Detection Kit is used to qualitatively detect mycoplasma contamination in cell culture. Over 95% of mycoplasma contamination in cell culture is caused by Mycoplasma fermentans, M. orale, M. pirum, M. hyorhinis, M. hominis, M. salivariu, M. arginine, and Acholeplasma laidlawii. Live mycoplasmas produce kinases not expressed in eukaryotic cells that convert ADP to ATP. The MYCO-LTM Luciferase Mycoplasma Detection Kit provides ADP and other substrates required by mycoplasma kinases, enabling the conversion of ADP to ATP. Luciferase-mediated ATP detection allows confirmation of mycoplasma contamination in cell culture. This kit rapidly detects these eight and many other common live mycoplasmas. Product Features: High Sensitivity: The MYCO-L™ Luciferase Mycoplasma Detection Kit has comparable sensitivity to nested PCR for mycoplasma detection. Convenient Protocol for Routine Monitoring of Mycoplasma Contamination in Cell Cultures: The protocol is simple and easy to use, with results available within 30 minutes. Detection of live mycoplasma contamination is particularly suitable for continuously monitoring the effectiveness of mycoplasma removal in cells. Residual mycoplasma DNA after mycoplasma removal can interfere with nested PCR and qPCR detection, resulting in false-positive results. The MYCO-L™ Luciferase Mycoplasma Detection Kit avoids these false-positive results by detecting live mycoplasmas.
Product composition:
| Specifications | Components | | 10T | Luciferase Mycoplasma Detection Reagent 0.5mL; Luciferase Mycoplasma Detection Substrate 0.1mL | | 25T | 50T | Luciferase Mycoplasma Detection Reagent 2.5mL; Luciferase Mycoplasma Detection Substrate 0.5mL |
| | General Notes | 1. Aliquot and Storage: Aliquot and store the reagents according to the instructions at -20°C to ensure stability.
2. Freeze-thawing two times of reagents stored at -20°C in the dark will not affect the test results.
3. Samples should be collected from cell cultures in the logarithmic growth phase for testing. Newly passaged cells should be sampled and tested 24-48 hours later.
4. The luciferase reaction is sensitive to temperature fluctuations. The reagents and test samples should be equilibrated to room temperature (22-25°C). The temperature should be maintained constant (±1°C) during the test. | | Storage Temp. | Store at -20℃ away from light, valid for 12 months. Transported by dry ice |
bio-equip.cn
AntBio is a biotechnology group company dedicated to serving life sciences, aiming to help scientists accelerate research and improve work efficiency. AntBio provides comprehensive and high-quality reagent tools for basic research, drug development, and diagnosis, including research grade antibodies, proteins, biochemical reagents, and assay kits. These research tools are widely used in different segments of life science research. The group company currently consists of three brands, Absin, Starter-Bio and UA-Bio.
|
中文
