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I have read that you do not offer a material in the kit to use as a standard in the afore mentioned MAOB assay. Could you please clarify where you attained the Hepg2 cells you did use in the assay? Were they from ATCC? If so what catalogue #? Additionally, I read that pargyline was use to stimulate the cells prior to using them as materials for the standard curve. Could you share that procedure? Did you use the same materials for controls? Do you have any historical data for intra or inter-assay precision? Thank you and have a great day.
Asked on Aug 22 2011
Thank you for contacting us! This assay is an immunoassay, a recombinant or purified standard would not be necessarily comparable to a native sample for activity so we don't offer it. For example an MAOB recombinant protein is available from Abcam ab82944 however it is not classified for functional studies funcS therefore we are not sure if it will work. The best control is MAOB isolated from untreated cells lysate. HepG2 cells are available from ATCC http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/tabid/452/Default.aspx?ATCCNum=HB-8065&Template=cellBiology MAOB is present in many human cell types we also have data, for example, with SHSY5Y cells. lastly, the data shown were in vitro (in-well), several exposure of drugs(including pargyline) in a dilution series to the isolated enzymes from normal untreated cells. The cells could be stimulated first then the enzyme isolated this may work -however we have not done this. The issue that may be faced here is that the extraction and dilution of the cell lysate into the assay buffer would change the local concentration of the pargyline thereby altering its effects on activity. I hope this information will be helpful. Should you have any other question please do not hesitate to ask.
Answered on Aug 22 2011