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Indirect Detection using IRDye® Labeled Streptavidin

Glycoprotein Detection with the Odyssey® Infrared Imaging System

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Description:

Biotinylated lectins are commercially-available from several vendors and can be used in combination with IRDye streptavidin to detect glycoproteins.

Detection Cartoon:

Slide 15

Data:

Slide 16

Transferrin contains N-linked glycoprotein structures and was separated on two reducing and denaturing 10% Bis-Tris gels. One gel was probed with biotinylated ConA and detected with IRDye 800CW streptavidin. The positive control was HRP (Horseradish Peroxidase).

Slide 17

A second gel was transferred to nitrocellulose, probed with biotinylated ConA and detected with IRDye® 800CW streptavidin. Both the gel and the membrane were imaged on the Odyssey and showed signal with the lectin, ConA. This confirmed the presence of mannose residues, indicative of N-linked glycan structures. The positive control was HRP (Horseradish Peroxidase).



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