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Invitrogen
Description: The mouse IgM is useful as an isotype control immunoglobulin. The immunoglobulin was generated by immunization with lipopolysaccharide (LPS). The immunoglobulin is reactive with LPS.
Applications Reported: eFluor® 660 Mouse IgM Isotype Control has been reported for use in flow cytometric analysis and immunohistochemical staining.
Applications Tested: eFluor® 660 Mouse IgM Isotype Control has been tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at the same concentration as the experimental antibody.
eFluor® 660 is a replacement for Alexa Fluor® 647. eFluor® 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome.
Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser.
Filtration: 0.2 µm post-manufacturing filtered.
IgM (Immunoglobulin M) is expressed intracellularly during the early stages of B lymphopoiesis, and then on the surface of more mature B cells in the bone marrow and peripheral B cells. The isotype of a primary antibody and its application 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 also 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.
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