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The isotype of a primary antibody and the application it is being used in can result in background staining. Primary antibody background noise can be caused by binding to Fc receptors on target cells; by non-specific interactions with cellular proteins, carbohydrates, and lipids; or by cell autofluorescence. Isotype control antibodies can act as negative controls to help differentiate non-specific background signal from specific antibody signal because they have no relevant specificity to a target antigen. While isotype controls are most commonly used in flow cytometry, they are useful in other applications such as chromatin immunoprecipitation (ChIP), immunohistochemistry, and gel shifts. Isotype controls should match with the primary antibody species and isotype so that the level of specific staining by the primary antibody may be accurately determined. If using directly labeled primary antibodies, the isotype control works best if conjugated with the same label as the test antibody.: This hamster IgG fraction was purified by affinity chromatography from normal Golden Syrian hamster serum.Applications Reported: This isotype control antibody has been reported for use in flow cytometric analysis.Applications Tested: This isotype control antibody has been tested by flow cytometric analysis of mouse splenocytes and normal human peripheral blood cells. It should be used at the same concentration as the experimental antibody.PerCP-eFluor™ 710 can be used in place of PE-Cy5, PE-Cy5.5 or PerCP-Cy5.5. PerCP-eFluor™ 710 emits at 710 nm and is excited with the blue laser (488 nm). Please make sure that your instrument is capable of detecting this fluorochrome. For a filter configuration, we recommend using the 685 LP dichroic mirror and 710/40 band pass filter, however the 695/40 band pass filter is an acceptable alternative. Our testing indicates that PerCP-eFluor™ 710 conjugated antibodies are stable when stained samples are exposed to freshly prepared 2% formaldehyde overnight at 4°C, but please evaluate for alternative fixation protocols.Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.Filtration: 0.2μm post-manufacturing filtered.
