The 3D7 antibody reacts with CD131, the 120 kD common β chain (βc) which is shared with the receptor complexes for human granulocyte-macrophage colony stimulating factor (GM-CSFR), interleukin-3 (IL-3R) and interleukin-5 (IL-5R). Together with the α subunit of either the IL-3R (IL-3Rα), IL-5R (IL-5Rα), or GM- CSFR (GM-CSFRα), the common β chain forms high-affinity, signaling receptors for human IL-3, IL-5 and GM-CSF, respectively. Cell surface βc are expressed by a variety of different cell types including hematopoietic progenitor cells derived from pluripotent stem cells, monocytes, neutrophils, eosinophils, basophils, endothelial cells, fibroblasts, and Langerhans cells. The immunogen used to generate this hybridoma was cells co-transfected with expression vectors which contained cDNA for the human IL-3 α and β chains.
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.