The MTS510 antibody reacts with the molecular complex of Toll-Like Receptor 4 and MD-2 (TLR4-MD-2) which is expressed on LPS responsive macrophages. TLR4, a member of the Toll-Like Receptor Family, has been renamed as CD284 and identified to be the transmembrane signal-transducing portion of the receptor for LPS. TLR4 associates on the cell surface with CD14 and MD-2, a 0.7 kDa molecule which is anchored to the membrane via its physical association with TLR4. The association of MD-2 with TLR4 is required for recognition of LPS and the anti-mitotic compound Taxol, which mimics the action of LPS on mouse cells. MTS510 mAb detects TLR4-MD-2 on the surface of thioglycollate-elicited macrophages from all mouse strains tested (ie, BALB/c, C57BL/6, C3H/HeJ, C3H/HeN, and DBA/1), including the C3H/HeJ strain which expresses an LPS-resistant mutant TLR4. Expression of TLR4-MD-2 is down-regulated on peritoneal macrophages after exposure to LPS, correlating with the occurrence of LPS tolerance. TLR4-MD-2 is not detected on splenocytes or thymocytes.
The antibody was conjugated to BD Horizon™ BV711 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 711-nm. BD Horizon BV711 can be excited by the violet laser and detected in a filter used to detect Cy™5.5 / Alexa Fluor® 700-like dyes (eg, 712/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there may be moderate spillover into the Alexa Fluor® 700 and PerCP-Cy5.5 detectors. However, the spillover can be corrected through compensation as with any other dye combination.