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UA-Glo Luminescent Mycoplasma Detection Kit

UA-Glo Luminescent Mycoplasma Detection Kit

Catalog Number: UA070102 Brand: UA BIOSCIENCE
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Regular price $316 USD
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Product Details

Product Specification


Synonyms Mycoplasma luciferase detection kits
Stability & Storage

Dry ice transport. -20℃ storage is protected from light, and the validity period is 12 months

Background

The UA-Glo Mycoplasma Detection Kit is used for the qualitative detection of mycoplasma contamination in cell cultures. Over 95% of mycoplasma contamination in cell cultures is caused by Mycoplasma fermentans, M. orale, M. pirum, M. hyorhinis, M. hominis, M. salivarius, M. arginine, and Acholeplasma laidlawii. Live mycoplasmas produce kinases that are not expressed by eukaryotic cells, which can convert ADP into ATP. The Mycoplasma Detection Kit provides ADP and other substrates required for mycoplasma kinases to convert ADP into ATP, and then detects the generated ATP via a luciferase luminescence method to confirm whether there is mycoplasma contamination in the cell culture. This kit can quickly detect the eight aforementioned species as well as many other common live mycoplasma strains.

Components

Size

component

10T

Mycoplasma luciferase detection reagent 0.5ml

Mycoplasma luciferase detection reagent 0.1mL

25T

Mycoplasma luciferase detection reagent 1.25ml

Mycoplasma luciferase detection reagent 0.25mL

50T

Mycoplasma luciferase detection reagent 2.5ml

Mycoplasma luciferase detection reagent 0.5mL


Protocol

1. Sample Preparation for Cell Culture Medium to be Tested

  1. Take 1 mL of cell culture medium from cells in the logarithmic growth phase.

  2. Centrifuge the cell culture medium at room temperature (200 g) for 5 minutes. Transfer approximately 800 μL of the supernatant to a new centrifuge tube, making sure not to include any cell debris at the bottom.

  3. Use the prepared cell culture medium for the mycoplasma detection with luciferase assay.

2. Luciferase Mycoplasma Detection

  1. Add 50 μL of the cell culture medium sample to a white opaque 96-well plate.

  2. Equilibrate the luciferase mycoplasma detection reagent and substrate to room temperature (22°C-25°C), and mix gently.

  3. Add 50 μL of the luciferase mycoplasma detection reagent to the 50 μL of the cell culture medium sample.

  4. Mix by gentle shaking for 10 seconds, incubate at room temperature for 5 minutes, and then read the fluorescence value (Reading 1).

  5. Add 10 μL of the luciferase mycoplasma detection substrate.

  6. Mix by gentle shaking for 10 seconds, incubate at room temperature for 10 minutes, and then read the fluorescence value (Reading 2).

3. Results
The ratio of Reading 2 to Reading 1 is used to determine if mycoplasma contamination is present.

 

The ratio of reading 2 to reading 1

Results Explanation and Recommendations

< 1

No Mycoplasma contamination

1-1.2

Possible Mycoplasma contamination. The cells were isolated and tested again after culture for 24-48hr.

> 1.2

Mycoplasma contamination



Guidelines

  • Aliquot Storage: Aliquot and store the reagents according to the instructions at -20°C to ensure reagent stability.

  • Freeze-Thaw Stability: Reagents stored at -20°C, protected from light, can be freeze-thawed twice without affecting the test results.

  • Sample Collection: Collect samples from cells in the logarithmic growth phase for testing. Newly passaged cells should be tested 24-48 hours after subculture.

  • Luciferase Reaction Sensitivity: The luciferase reaction is sensitive to temperature changes. Reagents and samples should be equilibrated to room temperature (22°C-25°C), and the temperature should remain constant (±1°C) throughout the test.

  • For Research Use Only: This product is intended for research purposes only.

Picture

Bioactivity

细胞UA-Glo荧光素酶支原体检测数据

阳性样品比值为4.15和6.68,该细胞存在支原体污染。其他培养基比值均<1, 提示不存在支原体污染。

UA试剂盒灵敏度优于L品牌:加入检测试剂和底物的相应读数1/2的荧光值(RLU)和读数2和1的比值。

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