The monoclonal antibody 55 recognizes lipoteichoic acid (LTA).
LTA, a glycerol phosphate surface polymer, is a component of the envelope of Gram-positive bacteria. LTA is anchored via its glycolipids to the membrane and carries a polysaccharide chain extending into the peptidoglycan layer of the cell wall. LTA is released spontaneously into the culture medium during growth of gram-positive bacteria. LTA functions as an immune activator with characteristics very similar to lipopolysaccharide (LPS) from Gram-negative bacteria. LTA binds to CD14 and triggers activation predominantly via Toll-like receptor 2. Although LTA is internalized and traffics to the Golgi, the cellular activation in response to LTA occurs at the cell surface.
Flow cytometry, Frozen sections, Immuno assays, Immuno fluorescence, Western blot
IA: For detection, 1.2 μg/ml antibody in PBS was added for 1 hr at 37 °C on LTA coated plates (Ref. 2,3,8).
IF: 60’in PBS/0.02%BSA/0.02% saponin (ref 4)
W: A reduced or native sample treatment and run on 15% SDS-Page. Blot was incubated o/n at 4°C with a 1/1000 dilution .The band size is ~17 kDa (Ref.6,7).
Dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.
Culture medium of Gram-positive bacteria.
Culture medium of eukaryotic cells