Whole cell lysate (20 μl) was combined with 80 μl of 0.05 N H2SO4 (hydrolysis reagent) and incubated at 80°C for 60 min. Samples were briefly centrifuged at 14,000 revolution/min (16,000 g), after which 20 μl of 1 M NaOH (neutralization reagent)...
To confirm if both α2,3 and α2,6 sialic acids facilitate AAV1 and AAV6 transduction, we carried out a lectin competition assay on these three cell lines (Fig. 5A to C). In competitive binding assays, 10 mm free synthetic sialic acid (Sigma) was...
We stained cell lines with lectins that have been used to recognize the following three different epitopes: (i) WGA recognizes all sialic acids, (ii) MAA recognizes α2,3 sialic acid, and (iii) SNA recognizes α2,6 sialic acid. In this report, we...
The data from these assays were complemented by a glycan array analysis which determined that AAV1 efficiently binds to NeuAcα2-3GalNAcβ1-4GlcNAc as well as glycoproteins containing NeuAcα2-3/α2-6Galβ1-4GlcNAc N-linked glycans. Sialic acids are...
Finally, is there a threshold of sialic acid loss that must be attained before barrier disruption occurs? More specifically, treatment of endothelial monolayers with neuraminidases leads to endothelial barrier disruption. However, the fact that...
Both mutants were grown on 2,7-anhydro-Neu5Ac (orange), Neu5Ac (blue), glucose (red), or M9 medium alone (black) in 200-µl microtiter plates. All strains were grown on 2,7-anhydro-Neu5Ac (orange), Neu5Ac (blue), glucose (red), or M9 medium alone...
Which application/end-user or product type may seek incremental growth prospects? AAV6 binding appears to be less affected by neuraminidase compared with AAV1 binding, suggesting that AAV6 may also bind to moieties other than sialic acid on the...
Creative Enzymes provides high-quality creatinase (EC 3.5.3.3), creatininase (EC 3.5.2.10), and sarcosine oxidase (EC 1.5.3.1), which are raw materials used in the production of creatinine assay kits. The viruses were removed by washing the...
For experiments, PC-12 cells were collected via 0.25% trypsin (Gibco) and seeded for culturing in collagen IV (Sigma-Aldrich) coated 4-chamber slides (Nunc Lab-Tek, Merck). PC-12 cells grew as small clusters in suspension and were maintained in...
Background was washed out with deionized water and gel was scanned with the default instrument settings for analysis (Lexmark CX727 scanner). Inhibition of glycolipid synthesis was carried out as described previously (36) with some...
