Lonza
MycoAlert(R) Mycoplasma Detection Kit
LT07-118
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My project involves analysis of the physiological role of the endocytic recycling regulator EHD4 in the kidney. EHD4 is expressed in the inner medullary collecting duct (IMCD) of the renal nephron, but its function here is unknown. The collecting duct is an important site that regulates water and salt homeostasis of the body. Mice lacking EHD4 develop phenotypes of dysregulated water and salt homeostasis. In order to understand the mechanism of how EHD4 works in the kidney, our lab uses a mouse collecting duct cell line. While working with cell lines, it is important to make sure that cells do not get contaminated with bacteria and mycoplasma. This kit allows for a routine test to detect mycoplasma contamination in cell culture.
Rapid, efficient mycoplasma detection assay
1ml Media from cell grown for 2 days in T-75 flask
The media was spun down at 200g for 5 mins and the supernatant was used. Make sure that the media is clear and does not contain any cells. Using a white plate, 25ul of media sample was loaded on one well and the assay buffer was loaded on another well as a negative control. 25ul of Assay Reagent was added (this is the minimum amount that gives reading). Wait for 5 min. Measure luminescence(Read A). Then add 25ul of substrate to the sample and control and wait 10 mins. Measure luminescence (Read B). Divide Read B by Read A to produce ratio.
MycoAlert Reagent and Substrate needs to be reconstituted with the Assay Buffer and kept at -20 for longer storage.
Ratio less than 0.9 means negative for mycoplasma. Ratio above 0.9 but less than 1.2 requires cells to be quarantined and retested in 24h. Ratio above 1.2 means positive for mycoplasma contamination.
None
Although the protocol says you to use 100ul of each reagent, you can actually use 25ul. So a 10 test kit should last you longer.
Expensive
Easy test for mycoplasma contamination.
Product Name: MycoAlert™ Mycoplasma Detection KitSupplier Page