. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. ,I. II Distribution of molluscan GAG (Fraction C) on ion-exchange chromatography and on cellulose acetate electrophoresis NaCl molarity Gills Mantle Ion exchange M M M 16% 38% 46% 19% 45% 33% Electrophoresis Migrating as: Heparan sulfate Heparin 55% 45% 79% 21% [35S] sulfate alone, or with up to 50 nCi/m\ radiolabeled glucosamine, 5 ^Ci/ml. The tissues were slowly shaken in a water bath at 25°C for 6 or 24 h and then washed with cold medium and ground in a Potter-Elvehjem tissue grinder in the presence of
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. ,I. II Distribution of molluscan GAG (Fraction C) on ion-exchange chromatography and on cellulose acetate electrophoresis NaCl molarity Gills Mantle Ion exchange M M M 16% 38% 46% 19% 45% 33% Electrophoresis Migrating as: Heparan sulfate Heparin 55% 45% 79% 21% [35S] sulfate alone, or with up to 50 nCi/m\ radiolabeled glucosamine, 5 ^Ci/ml. The tissues were slowly shaken in a water bath at 25°C for 6 or 24 h and then washed with cold medium and ground in a Potter-Elvehjem tissue grinder in the presence of 1% Triton X-100 in 4 M urea containing the protease inhibitors. The ground material was added to a DEAE-Sephacel column and washed ex- tensively with the above solution in AI NaCl. The labeled material was then eluted with the 1% Triton, 4 M Kav Figure 2. Sepharose CL-4B. Chromatography of material isolated from the peaks shown in Figure 1. Top: labeled material from gills in peak B before and after treatment with pronase. Solid line, untreated material; shaded line, pronase digested material. Bottom: labeled material from mantle in peaks A, B. C shown in Figure I after alkaline borohydride treatment. Material from peak A, shaded line; peak B, and peak C, solid line. Note that compared to Figure 1. material in peaks A and B shifted to a KjV of whereas material in peak C (with a Kav of ) showed no further shift. trophoresis in AI formic acid at pH Scintillation counting was performed on a Packard Tri Carb 1500 using Opti Fluor scintillation cocktails. The presence of proteoglycans was determined by al- kaline degradation with borotritide in N NaOH for 16 h at 4°C or in WNaOH for 16 h at room temper- ature. In this connection, the term 'free chains' (vs. pro- teoglycans) is used here as denning GAGs that show no change in molecular weight upon alkaline treatment; they may contain a small peptide, however. Organ culture Gills and mantle
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
