scholarly journals Fractionation of Buffalo Milk Casein by Acrylamide Gel Electrophoresis and DEAE Cellulose Column Chromatography

1973 ◽  
Vol 56 (1) ◽  
pp. 61-65 ◽  
Author(s):  
Taro Nagasawa ◽  
Isao Kiyosawa ◽  
Kunisuke Kuwahara ◽  
N.C. Ganguli
Keyword(s):  
Column Chromatography ◽  
Gel Electrophoresis ◽  
Deae Cellulose ◽  
Buffalo Milk ◽  
Deae Cellulose Column ◽  
Milk Casein ◽  
Cellulose Column

scholarly journals Purification and chemical characterization of human hexosaminidases A and B

1976 ◽  
Vol 159 (3) ◽  
pp. 535-539 ◽  
Author(s):  
J E S. Lee ◽  
A Yoshida
Keyword(s):  
Column Chromatography ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Polyacrylamide Gel ◽  
Gel Chromatography ◽  
Deae Cellulose Column ◽  
Hexosaminidase A ◽  
Cellulose Column

N-Acetyl-β-hexosaminidases A and B were purified to homogeneity from human placenta. In the initial step of purification, the enzymes were adsorbed on concanavalin A-Sepharose 4B and eluted from the column with α-methyl D-mannosides. Subsequent purification steps included DEAE-cellulose column chromatography, QAE-Sephadex [diethyl-(2-hydroxypropyl)aminoethyl-Sephadex] column chromatography, Sephadex G-200 gel filtration and preparative disc polyacrylamide-gel electrophoresis, followed by another QAE-Sephadex chromatography for the hexosaminidase A preparation, and DEAE-cellulose column chromatography, calcium phosphate gel chromatography, Sephadex G-200 gel filtration, QAE-Sephadex chromatography and CM-cellulose chromatography for the hexosaminidase B preparation. The purified preparations, particularly hexosaminidase A, had significantly higher specific enzyme activities than previously reported. The preparations moved on polyacrylamide-gel electrophoresis as single protein bands, which also stained for enzyme activity. Sedimentation-equilibrium centrifugation indicated homogenous dispersion of the enzymes, and the molecular weight was estimated as about 110000 for both enzymes. Complete amino acid and carbohydrate compositions of the two isoenzymes were determined, and, in contrast with previous suggestions, no sialic acid was found in the enzymes.

A simple assay for galactokinase using deae-cellulose column chromatography

1977 ◽  
Vol 74 (1) ◽  
pp. 1-5 ◽  
Author(s):  
Y.S. Shin-Buehring ◽  
M. Osang ◽  
R. Ziegler ◽  
J. Schaub
Keyword(s):  
Column Chromatography ◽  
Deae Cellulose ◽  
Deae Cellulose Column ◽  
Cellulose Column

scholarly journals Studies on ox brain aminopeptides

1967 ◽  
Vol 105 (2) ◽  
pp. 641-646 ◽  
Author(s):  
A. S. Brecher ◽  
R. E. Sobel
Keyword(s):  
Ammonium Sulphate ◽  
Column Chromatography ◽  
Deae Cellulose ◽  
Deae Cellulose Column ◽  
Cellulose Column ◽  
Inhibited Enzyme

1. Mn2+-inhibited and Mn2+-activated aminopeptidases have been observed in ox brain and separated from one another by DEAE-cellulose column chromatography. 2. The Mn2+-inhibited enzyme has been purified 36-fold; it exhibits a specificity for tripeptide substrates, whereas the Mn2+-activated aminopeptidase cleaves dipeptides as well as tripeptides. 3. Ammonium sulphate treatment generates a Mn2+-stimulated aminopeptidase that is stable to dialysis against EDTA and water, in contrast with an endogenous Mn2+-activated preparation that is irreversibly denatured by such dialysis against EDTA and water.

scholarly journals Purification and characterization of biotin-binding protein II from chicken oocytes

1988 ◽  
Vol 256 (3) ◽  
pp. 797-805 ◽  
Author(s):  
L Bush ◽  
T J McGahan ◽  
H B White
Keyword(s):  
Amino Acid ◽  
Column Chromatography ◽  
Gel Electrophoresis ◽  
Binding Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Deae Cellulose ◽  
Polyacrylamide Gel ◽  
Chicken Egg ◽  
Biotin Binding ◽  
Cellulose Column

BBP-II, the major biotin-binding protein from chicken oocytes, was purified 12,000-fold with a 22% yield. The purification procedure includes butan-1-ol extraction of yolk lipids, phosphocellulose chromatography of the water-soluble proteins, DEAE-cellulose chromatography at pH 7.4 and hydroxyapatite column chromatography. Final purification was obtained by using a second DEAE-cellulose column chromatography at pH 6.0. BBP-I activity separated from BBP-II activity during elution from the first DEAE-cellulose column. Purified BBP-II was homogeneous on both polyacrylamide-gel electrophoresis and SDS/polyacrylamide-gel electrophoresis under conditions that would detect a 1% impurity. The subunit Mr determined from SDS/polyacrylamide-gel electrophoresis was 18,200 (72,600 for tetramer), which compares favourably with an Mr value of 17,300 (69,100) calculated from the amino acid analysis. A single precipitin line formed when rabbit antiserum to the protein was directed against a crude chicken egg-yolk sample. BBP-II purified by this procedure lacked carbohydrate and phosphate, was stable indefinitely when frozen, and was quite stable at room temperature. The N-terminal amino acid sequence showed polymorphism at three positions in the first 23 residues and was about 45% identical with the N-terminal 22 residues of avidin. Antiserum to BBP-II cross-reacted with BBP-I and similar proteins in the yolk of eggs from various birds and alligator as judged by immunodiffusion and enzyme-linked immunosorbent assays. No cross-reaction was observed with chicken egg-white by either of these methods.

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