Abstract:
:Flow cytometry was used to demonstrate the presence of beta-lactoglobulin (betaLG) receptors on living murine hybridoma MARK-3 cells using a fluorescein isothiocyanate-betaLG conjugate (FITC-betaLG: molar ratio of 5:1). A site occupation curve was produced using a shift in the mean channel fluorescence at various concentrations of FITC-betaLG. The binding of labelled ligand was concentration dependent and was inhibited by unlabelled betaLG. The on-rate constant was 3.2x10(2) M(-1) min(-1) and the off-rate constant was 0.002 min(-1). Scatchard plot analysis gave a dissociation constant (K(d)) of 44+/-21x10(-7) and 39+/-24x10(-5) M (n=3). Flow cytometry indicated that at least 15% of the FITC-betaLG were internalized for 5 min and that internalization was temperature- and time-dependent. The internalization was confirmed by 3-D fluorescence microscopy (CELLScan system).
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Palupi NS,Franck P,Guimont C,Linden G,Dumas D,Stoltz J,Nabet P,Belleville-Nabet F,Dousset Bdoi
10.1016/s0168-1656(00)00210-8subject
Has Abstractpub_date
2000-03-10 00:00:00pages
171-84issue
2eissn
0168-1656issn
1873-4863pii
S0168-1656(00)00210-8journal_volume
78pub_type
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