Ws data station

The glycolipid AT-X was treated with 3 m HCl in CH₃OH (Supelco) overnight at 80 °C to both release the fatty acyl groups from AT-X and form their methyl esters. The sample was then dried and dissolved in 50 μl of N,O-bis(trimethylsilyl) trifluoroacetamide (Sigma-Aldrich) and heated at 60 °C for 10 min prior to injection for GC/MS to form the trimethylsilyl ethers of any hydroxyl groups. Samples were injected directly from the silylating reagent. Analyses were carried out using a CP 3800 gas chromatograph (Varian) equipped with an MS320 mass spectrometer in the electron impact mode and scanning from m/z 50 to 800 over 0.5 s. Helium was used as the carrier gas with a flow rate of 1 ml/min. The samples were run on a DB 5 column (10 m × 0.20-mm inner diameter). The injector (splitless mode) was set for 250 °C. The oven temperature was held at 50 °C for 1 min, programmed at 30 °C/min to 130 °C, and then programmed at 10 °C/min to 330 °C, followed by a 10-min hold. The data analyses were carried out on a Varian WS data station.

Most of the reagents were bought from Sigma-Aldrich and used without further purification. The silylation agent, HTP-trisil, was bought from Thermo-Fisher Scientific. Solvents were purchased from Fisher Scientific. Uniformly labeled ¹³C₅-D-arabinose was purchased from Cambridge Isotope Laboratories, Inc. Octyl sepharose CL 4B was obtained from GE Healthcare. GC/MS was recorded on CP 3800 gas chromatograph (Varian) equipped with an MS320 mass spectrometer. Helium was used as the carrier gas with a flow rate of 1 ml/min. Argon was used as the collision gas in MS/MS experiments and methane was used as the chemical ionization gas for the negative chemical ionization analyses. The samples were analyzed on an Agilent J&W capillary column VF-5ms column (30 m × 0.25 mm i.d.). The data analyses were carried out on a Varian WS data station. NMR spectroscopy was carried out on Innova 400 (Varian) at the Central Instrument Facility, CSU. Healthy nonendemic urine (NEU) samples were collected from willing co-worker-volunteers in CSU. The primary antibody CS-35, used in the dot blot assay was developed in our laboratory and the detection antibody anti-mouse IgG alkaline phosphatase was purchased from Sigma-Aldrich