Bovine beta-lactoglobulin receptors on transformed mammalian cells (hybridomas MARK-3): characterization by flow cytometry.

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 Biotechnol

journal_title

Journal of biotechnology

authors

Palupi NS,Franck P,Guimont C,Linden G,Dumas D,Stoltz J,Nabet P,Belleville-Nabet F,Dousset B

doi

10.1016/s0168-1656(00)00210-8

subject

Has Abstract

pub_date

2000-03-10 00:00:00

pages

171-84

issue

2

eissn

0168-1656

issn

1873-4863

pii

S0168-1656(00)00210-8

journal_volume

78

pub_type

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