>>> Maciej Simm <simmmmer@yahoo.com> 03/20/01 09:05PM >>> >> >> Sometimes I read people's materials and methods "irrelevant >>isotype-matched fluorescent antibody was used.." what exactly does >>that mean? Maciej, This refers to a control that is isotype (class and subclass) matched with the specific antibody you're interested, but is "irrelevant" to the target . . . i.e., there's no way the control's targeted antigen could be there. Example: BD's Mouse-anti-Human CD45 is types as a IgG, subclassed as an IgG1. The irrelevant negative IgG1 is made in mouse to a keyhole limpet hemacyanine antigen. It's the same isotype as the CD45 ab, but there's no chance of finding the hemacyanine antigen on your leukocytes (unless, of course, you're a keyhole limpet). Actually, I searched the BD site, and found this < http://www.bdfacs.com/source_book/pdf/23-1348-11.pdf > . . . just what I'm talking about. Re. these negative controls . . . 1) you also need to match conjugate colors, and 2) there are flowists who are of the opinion that these controls are unnecessary . . . I'm not one of them. MAK. -- Mark A. KuKuruga, Managing Director University of Michigan Flow Core 7416 CCGC 0946 (734) 647-3216, fax (734) 936-7376 kukuru@umich.edu
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