PlasmoTest™ provides a simple, rapid and reliable assay for the visual detection of mycoplasma contamination in cell cultures. This assay is the first to utilize cells to signal the presence of mycoplasma.
Benefits
• Simple - Requires only basic cell culture knowledge. No need for specific lab equipment. Results are easily determined with the naked eye or quantified with a spectrophotometer. • Rapid - Hands-on time less than 1 hour. Gives results after overnight incubation. • Versatile - Detects all Mycoplasma and Acholeplasma species known to infect cell cultures, as well as other cell culture contaminants such as bacteria. • Sensitive - Detects 5.102-5.105 cfu/ml mycoplasmas. No false positive: a positive result indicates the presence of a cell culture contaminant. • Complete - Contains the Mycoplasma sensor cells and all the reagents needed to perform the assay, including positive and negative controls. Up to 500 samples can be tested with the kit. To perform further assays, only the reagents need to be reordered.
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Principle
PlasmoTest™ features two major constituents: the Mycoplasma sensor cells and the HEK-Blue™ Detection medium. The Mycoplasma sensor cells detect the presence of mycoplasmas leading to a color change of the HEK-Blue™ Detection medium. The Mycoplasma sensor cells recognize mycoplasmas through Toll-Like Receptor 2 (TLR2), a pathogen recognition receptor. In the presence of mycoplasmas, TLR2 initiates a signaling cascade leading to the activation of NF-κB and other transcription factors. These transcription factors induce the secretion of SEAP (secreted embryonic alkaline phosphatase) in the supernatant which is readily detected by the purple/blue coloration of the HEK-Blue™ Detection medium.
PlasmoTest™ Principle
Key Features
HEK-Blue™-2 cells, the Mycoplasma sensor cells, are engineered HEK293 cells. These cells stably express TLR2 and multiple genes from the TLR2 pathway and coexpress an optimized SEAP reporter gene, placed under the control of a promoter inducible by the transcription factors NF-κB and AP-1.
HEK-Blue™ Selection is a solution that combines several selective antibiotics. These antibiotics guarantee the persistent expression of the various transgenes introduced in HEK-Blue™-2 cells. Furthermore, Normocin™ is included in the kit to protect HEK-Blue™-2 cells from any potential microbial contamination, whether caused by mycoplasmas, bacteria or fungi.
HEK-Blue™ Detection is a medium specifically designed for the detection of SEAP. It contains a color substrate that produces a purple/blue color following its hydrolysis by SEAP.
Important Licensing Information: These products are covered by a Limited Use License (See Terms and Conditions). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses.
PlasmoTest™ Procedure
1- Collect 500 μl cell culture supernatant and heat samples at 100°C for 15 mins. Add 50 μl of each sample and control in a well of a 96-well plate. 2- Prepare HEK-Blue™-2 cell suspension with HEK-Blue™ Detection medium. Add 200 μl (~50,000 cells) of cell suspension to each well containing the samples or controls. 3- Incubate the plate at 37°C in a CO2 incubator overnight (16-24h). 4- Detect the presence of Mycoplasma with the naked eye or by reading the OD at 620-655 nm.
Contents
PlasmoTest™ Kit includes: - HEK-Blue™-2 cells (3-5 x 106 cells) Cells are shipped on dry ice. Upon receipt, store HEK-Blue™-2 cells at -80°C. - HEK-Blue™-2 Selection (4 x 2 ml) - 250X Antibiotic mix - Normocin™ (4 x 1 ml) - HEK-Blue™ Detection (2 x 50 ml) - HEK-Blue water (120 ml) - PlasmoTest Controls (200 tests)
PlasmoTest Controls include a positive and a negative control (200 tests). Note: the positive control has changed for PlasmoTest™ kits sold after March 15, 2009. The former posiitve control is still available upon request.
Buy the PlasmoTest™ kit once then reorder only the reagents to perform further assays. The reagents can be purchased individually or together as the PlasmoTest™ Reagent Kit which includes the following: - HEK-Blue™ Selection (4 x 2 ml) - Antibiotic mix - Normocin™ (4 x 1 ml) - HEK-Blue™ Detection (2 pouches) - HEK-Blue™ water (2 x 60 ml) - Positive control - Negative control
Disclaimer: Our products are provided for research purpose only. Commercial applications may require licensing from third parties.
Note that the sequence of available ORFs provided by InvivoGen can differ from a given reference Genbank record due to genetic variations
and/or alternative splicing. Customers should verify that the version of a gene sold by InvivoGen is suitable for the customer needs.
