Description: 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 can be used at less than or equal to 0.125 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.
PE-eFluor® 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red®). Please make sure that your instrument is capable of detecting this fluorochome.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
Filtration: 0.2 µm post-manufacturing filtered.
MHC (major histocompatibility complex) class II molecules are transmembrane glycoproteins expressed on the surface of professional antigen-presenting cells, such as macrophages, dendritic cells and B cells. Before their exposition on the cell surface, the MHC class II molecules react with endocytosed exogenous antigens, which are then presented to the T cells. The antigen-binding grove between MHC class II alpha and beta chain is open at both ends and is 15-24 amino acid residues long.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: cell surface glycoprotein; H-2 class II histocompatibility antigen, A beta chain; H-2 class II histocompatibility antigen, I-E beta chain; histocompatibility 2, class II antigen A, beta 1; I-Ab; I-Ad; I-Aq; I-E beta MHC class II; I-Ed; I-Ek; major histocompatibility complex class II beta chain; major histocompatibility complex H-2E beta chain; MHC class 2; MHC class II antigen A beta; MHC class II H2-IA-beta-psi; MHC class II IE-b; MHC H2-IE-beta cell surface glycoprotein; response to metastatic cancers 1
Gene Aliases: Abeta; AI845868; H-2Ab; H-2Eb; H2-Ab; H2-Ab1; H2-iabeta; H2Eb; I-Abeta; Ia-2; Ia-4; Ia2; Ia4; IAb; Rmcs1
UniProt ID: (Mouse) P14483
Molecular Function: major histocompatibility complex protein