Journal:

Article Title: Glycosylation of the Major Polar Tube Protein of Encephalitozoon hellem , a Microsporidian Parasite That Infects Humans

doi: 10.1128/IAI.72.11.6341-6350.2004

Figure Lengend Snippet: Interaction of EhPTP1 with the lectin ConA. (A) ConA overlay. A DTT-solubilized polar tube preparation was used for SDS-PAGE, transferred to nitrocellulose, and incubated with labeled ConA in the presence and absence of α-methyl-mannopyranoside. ConA bound to a single band (lane 1), and this binding was inhibited by α-methyl-mannopyranoside (lane 2). (B) ConA affinity chromatography. Soluble E. hellem PTPs were loaded onto a ConA-Sepharose column, and specifically retained proteins were eluted with 0.2 M α-methyl-mannopyranoside-0.15 mM NaCl. The flowthrough of the column (e.g., soluble PTPs after loading on the column) and the specific eluate were analyzed by SDS-PAGE followed by immunoblot analysis with rabbit antibody to recEhPTP1. The specific eluate contained EhPTP1 (lane 1), while the flowthrough no longer contained EhPTP1 (lane 2).

Article Snippet: DTT-solubilized PTPs (200 μg) were subjected to ConA affinity chromatography (0.5 ml of ConA gel [EY Laboratories, Inc.]).

Techniques: SDS Page, Incubation, Labeling, Binding Assay, Affinity Chromatography, Western Blot

Journal:

Article Title: Glycosylation of the Major Polar Tube Protein of Encephalitozoon hellem , a Microsporidian Parasite That Infects Humans

doi: 10.1128/IAI.72.11.6341-6350.2004

Figure Lengend Snippet: PTP1 reactivity with ConA is seen in multiple microsporidia. Solubilized (2% DTT) polar tube preparations of five microsporidia species, G. americanus (Ga), E. hellem (Eh), E. cuniculi (Ec), E. intestinalis (Ei), and B. algerae (Ba), were used for a lectin overlay with labeled ConA. ConA reacted with a band in each polar tube preparation that had a size consistent with that seen on SDS-PAGE and immunoblot analysis with antibody to PTP1 (12-15).

Article Snippet: DTT-solubilized PTPs (200 μg) were subjected to ConA affinity chromatography (0.5 ml of ConA gel [EY Laboratories, Inc.]).

Techniques: Labeling, SDS Page, Western Blot