Figure 1. RTL1000 is highly purified, monomeric, biotinylated and biologically active.

A) Purified RTL1000 was analyzed by Size Exclusion Chromatography and detected by absorbance at 280nm. * Indicates elution of known molecular weight proteins (43, 29, 13.7, 6.5kD)with increasing retention volumes. Insert: RTL1000 sample was analyzed by SDS-polyacrylamide gel electrophoresis (left lane) and compared to MW standards (right lane). B) Samples of RTLs with or without β-mercaptoethanol, analyzed by SDS-polyacrylamide gel. C) Biotinylated RTL1000 and 100% biotinylated MBP standard were separated by SDS-PAGE, blotted and stained with HRP-conjugated streptavidin. D) Inhibition of the MOG-35-55-specific response of H2-1 T-cell hybridoma by RTL1000. H2-1 cells were pre-incubated with RTL1000, RTL340 or medium alone before their Ag-specific activation with DR*1501 APC pulsed with MOG-35-55 peptide compared to medium alone. CTLL proliferation (triplicate mean of ³H-Tdy uptake) differences between the samples pre-incubated with RTL1000 and medium alone were evaluated using two-tailed t-test. Data in A–D are representative of at least three independent experiments.