Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose, ProClin 300.
Expression System:
HEK293 cells
Formulierung:
Lyophilized powder
Sequenz:
N-terminal His-Tag, Ala141~His286 (NP_000425.1)
Anwendungsbeschreibung:
NEU (Sialidase-1) is an enzyme that catalyzes the removal of sialic acid (N-acetylneuraminic acid) moities from glycoproteins and glycolipids. In the lysosome, this enzyme is part of a heterotrimeric complex together with beta-galactosidase and cathepsin A (CTSA). Thus a binding ELISA assay was conducted to detect the interaction of recombinant human NEU and recombinant human CTSA. Briefly, NEU were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 µl were then transferred to CTSA-coated microtiter wells and incubated for 2h at 37C. Wells were washed with PBST and incubated for 1h with anti-NEU pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37C. Finally, add 50 µl stop solution to the wells and read at 450nm immediately. The binding activity of NEU and CTSA was in a dose dependent manner.
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