IgM Monoclonal Antibody (II/41), eFluor™ 450, eBioscience™, Invitrogen™

Description

Description: The II/41 monoclonal antibody reacts with the mu heavy chain of mouse IgM. It does not react with other classes of mouse immunoglobulin including IgD, IgG or IgA. IgM is expressed intracellularly, during early stages of B lymphopoiesis, and then on the surface of more mature B cells in the bone marrow and peripheral B cells. Fluorochrome conjugated II/41 can be used as a detection secondary for mouse IgM. Applications Reported: This II/41 antibody has been reported for use in flow cytometric analysis. Applications Tested: This II/41 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This can be used at less than or equal to 0.5 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. eFluor® 450 is an alternative to Pacific Blue®. eFluor® 450 emits at 445 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 405 nm; Emission: 445 nm; Laser: Violet 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.