The M5/114.15.2 monoclonal antibody reacts with the mouse major histocompatibility complex class II, both I-A and I-E subregion-encoded glycoproteins (I-A b, I-A d, I-A q, I-E d, I-E k, not I-A f, I-A k, or I-A s). It detects a polymorphic determinant present on B cells, monocytes, macrophages, dendritic cells, and activated T lymphocytes from mice carrying the H-2 b, H-2 d, H-2 q, H-2 p, H-2 r and H-2 u but not from mice carrying the H-2 s or H-2 f haplotypes. The M5/114 mAb is reported to inhibit I-A-restricted T cell responses of the H-2 b, H-2 d, H-2 q, H-2 u but not H-2 f, H-2 k, or H-2 s haplotypes.

Applications Reported: This M5/114.15.2 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This M5/114.15.2 antibody has been tested by flow cytometric analysis of mouse splenocytes. This may be used at less than or equal to 0.4 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.

NovaFluor dyes are not compatible with DNA intercalating viability dyes. Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.

Each NovaFluor conjugate or kit is shipped with CellBlox Blocking Buffer. Use this buffer whenever staining with NovaFluor conjugates, including single-color compensation controls using cells. Whenever possible, we recommend adding CellBlox Blocking Buffer to antibody cocktails/master mixes prior to combining with cells. Add 5 µL per sample (regardless of the number of NovaFluors in your panel) to use the antibody cocktail as intended. For single-color controls, use 5 µL of CellBlox Blocking Buffer per 100µL of cell sample containing 10^3 to 10^8 cells.

Excitation: 496 nm; Emission: 511 nm; Laser: 488 nm (Blue) Laser

NovaFluor conjugates are based on Phiton™ technology utilizing novel nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more