Lectin-sepharose

In a preliminary report, Ross et al. [40] used affinity chromatography to identify a putative bovine renal brush border Na /H exchanger. Brush border membranes were solubilized with Triton X-100 and chromatographed sequentially over lentil lectin Sepharose 4B and 5-(A-benzyl-iV-ethyl)amiloride coupled to epoxy-activated Sepharose 6B. The eluant contained 178- and 146-kDa proteins that were susceptible to Endo-F. Moreover, the eluants reacted on dot blot immunoassays with antisera to a 20-amino acid peptide of a human Na /H exchanger vide infra). The relationship between these proteins and the 66-kDa protein previously identified by the same investigators using amiloride photolabeling is presently unclear. 

Lentil lectin-Sepharose Detergent-soluble membrane proteins... 

Lentil Lectin Sepharose Wheat Germ Lectin Lectins Membrane proteins, glycoproteins, polysaccharides Pharmacia... 

A mixture of lentil lectin-reactive glycoproteins from pig lymphocyte-plasma membrane was isolated by lentil lectin-Sepharose chromatography of sodium deoxycholate-solubilized membranes.445 Eighty-seven percent of the protein applied (17% of hexose) passed through unretarded, and 13% of the applied protein (83% of hexose) was bound, and eluted with methyl a-D-glucopyranoside solution. Recovery was 95% of the material applied, in contrast to the recovery in similar experiments conducted on con A-Sepharose columns (80% recovery).850 The eluate from the lentil column, which contained at least ten glycoproteins, blocked lymphocyte transformation induced by lentil or kidney-bean lectins.445... 

Nachbar and coworkers investigated the composition of Ehrlich ascites, tumor-cell plasma membrane by using a series of lectin-Sepharose adsorbents (R. communis hemagglutinin, con A, wheat-germ agglutinin, and soybean agglutinin).868 The Ehrlich cells... 

Bovine viral diarrhea virus Crotolaria juncea lectin Sepharose 2B 207... 

Red blood cells bound to Concanavalin A - Sepharose or Lens cull nan s lectin - Sepharose was used in combination with immo-bilized glucose oxidase (E.C. 1.1.3.4) in the thermistor system. The red blood cells operated as oxygen supply in the reactor bed leading to better reaction conditions for the oxidase - thereby widening the practical concentration range for glucose analysis... 

D-Glucose oxidase and peroxidase, when adsorbed on to concanavalin A-Sepharose or to Lens culinaris lectin-Sepharose have been used in an analytical system for the estimation of D-glucose. ... 

Lentil lectin Sepharose 4B 4% agarose Lentil lectin 2mg/mL 3-10 45-165 Membrane proteins AP Biotech ... 

Wheat Germ Lectin Sepharose 6MB 6% agarose Wheat germ lectin 5mg/mL n.a. 200-300 Glycoproteins, polysaccharides, membrane proteins AP Biotech C- c 3 (fi O fi)... 

Sepharose-lectin column chromatography To purify glycoproteins or gly-copeptides that bind the particular lectin used. 

Membrane extracts from adult H. contortus were enriched 24-fold for cysteine protease activity by passage over a Thiol-Sepharose affinity column and the proteins obtained (abbreviated as TSBP) were clearly localized to the microvillar surface of the intestinal cells (Knox et al., 1995,1999). TSBP comprised a prominent 60 kDa protein and several minor bands between 35 and 45 kDa and 97 to 120 kDa (Fig. 13.2). Protease activity at 38, 52 and 70 kDa was attributable to cysteine proteases and at 70 and 88 kDa to serine/metalloproteases, as judged by inhibition analyses. Lectin-binding studies showed that most of the TSBPs were glycosylated. Expression library... 

Cuatrecasas (45). The Dolichos biflorus lectin was a gift from Dr. Marilynn Etzler (46). Ulex europeus and Bandeirea simplicifolia lectins were purified in our laboratory and iodinated with Nal25i (carrier-free) by Sepharose 4B-bound lactoperoxidase (47)and diazotized iodoaniline coupling (48) procedures, respectively. 

One approach to this problem would be to isolate the intact lectin-receptor molecules from the cell membranes and characterize them. Work in this direction has been initiated in our laboratory, and a method for the isolation of the peanut agglutinin receptor from membranes of neuraminidase-treated human erythrocytes on a column of peanut agglutinin-polyacryl-hydrazido-Sepharose has been developed (21). The amino acid composition, D-glucosamine ancHg-galac-tosamine content, and the electrophoretic mobility on polyacrylamide gel electrophoresis in sodium... 

An alternative approach involves insolubilized glycoproteins. Thus, an affinity system for the isolation of the lectin of red kidney-bean (Phaseolus vulgaris)149,154 involved thyroglobulin-Sepharose, and, for... 

Attempts to purify the potato lectin on ovomucoid- and N,N -diacetylchitobiose-substituted Sepharose failed the lectin was so strongly bound that it could not be removed from the adsorbent by displacement with urea.483 To circumvent this difficulty, Delmotte and coworkers483 employed p-aminobenzyl 2-acetamido-2-deoxy-l-thio-/3-D-glucoside-substituted Sepharose, and eluted the lectin with 0.1 M acetic acid (yield 58 mg from 128 g of the protein in potato-tuber extract). 

Cell-binding studies on human erythrocytes, several human, urinary-bladder, carcinoma cell-lines, and an osteogenic-sarcoma cellline have been conducted.189 After treatment with neuraminidase, 80% of human lymphocytes will bind the H. pomatia lectin.5728 Neuraminidase-treated lymphocytes can also be fractionated on Helix pomatia hemagglutinin coupled to Sepharose beads.572b... 

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