The LMM741 monoclonal antibody specifically recognizes CD114 which is also known as the Granulocyte-Colony Stimulating Factor Receptor (G-CSFR). CD114 is a ~150 kDa type I transmembrane glycoprotein that is encoded by CSF3R (colony stimulating factor 3 receptor) and belongs to the class 1 cytokine receptor family. CD114 is expressed on granulocytes, monocytes, dendritic cells, endothelial cells, platelets, placenta and myeloid leukemias and a variety of tumor cell lines. CD114 serves as the receptor for granulocyte colony stimulating factor (G-CSF) which plays a role in myeloid cell proliferation and differentiation. The immunogen used to generate this hybridoma was cells transfected with an expression vector containing a full-length cDNA encoding the human G-CSFR.
The antibody was conjugated to BD Horizon™ BUV496 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 496-nm. BD Horizon BUV496 can be excited by the ultraviolet laser (355 nm) and detected with a 515/30 nm filter with a 450LP. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into the channel detecting BD Horizon V500 or BV510 (eg, 525/40-nm filter). However, the spillover can be corrected through compensation as with any other dye combination.