The photoreactive carbohydrate probe, a novel method for detection of lectins.
One of the main difficulties in the research of lectins is the absence of an adequate technique for their specific and routine detection. Here, we present a photoreactive carbohydrate-probe which could help to overcome this problem. The probe was comopsed by joining four segments: (i) a carbohydrate moiety, (ii) the digoxigenin tag, (iii) the photoreactive cross-linker and (iv) the lysyl-lysine backbone. After incubation with lectins in the dark, the probe can be activated and cross-linked to the lectins by illumination. The result is a lectin with covalently incorporated digoxigenin tag. Such a labelled lectin can be easily identified using anti-digoxigenin antibodies in a Western blot technique. This method is of high specificity and sensitivity and enables direct detection of lectins in complex mixtures, even whole cell homogenates.
Lauc
G
G
Department of Biochemistry and Molecular Biology, University of Zagreb, Croatia.
Barisić
K
K
Zanić
T
T
Flögel
M
M
D002241:Carbohydrates / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry; D004076:Digoxigenin / Q000737:chemistry; D005947:Glucose / Q000737:chemistry; D037102:Lectins / Q000032:analysis*; D008239:Lysine / Q000737:chemistry; D010777:Photochemistry / Q000379:methods; D012680:Sensitivity and Specificity
D002241:Carbohydrates; D003432:Cross-Linking Reagents; D037102:Lectins; D005947:Glucose; D008239:Lysine; D004076:Digoxigenin