two step purification of a 66 kda-endoxylanase and a broad specificity endoglucanasexylanase from...
DESCRIPTION
BioseparationsTRANSCRIPT
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Separation Mode: Anion Exchange Chromatography
Instrumentation: Gradient HPLC system
Separation Device: Same CIM DEAE tube for both steps; Active bed: Outer diameter: 15 mm, Inner diameter:1.1 mm, Length: 45 mm, Volume: 8 mL
APP_001 BLACK PANTONE 320 CV SYNCOMP
BIA
Separatio
ns
CIM CONVECTIVE INTERACTION MEDIA
APPLICATION SHEETS
P001
First separation step shown in Figure A:
Sample: Non-concentrated periplasmic fraction(0.14 mg protein/mL) from Prevotella bryantii cultivationmedium; 1, 2, 3, 4, 5 and 6 are collected fractions
Applied Sample Volume: 60 mL
Mobile Phase: Buffer A: 20 mM Tris-HCl, pH 7.4;Buffer B: Buffer A + 1 M NaCl, pH 7.4
Conditions: Gradient: as shown in Figure A;Flow Rate: 7 mL/min; Detection: UV at 280 nm
Two Step Purification of a 66 kDa-Endoxylanase and a BroadSpecificity Endoglucanase/Xylanase from Prevotella bryantii B14on a CIM DEAE-8 Tube
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Figure A:
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CIM technology is covered by US patents 4889632, 4923610, 4952349 and 5972218. Other patents pending.
19992000 by BIA Separations d. o. o. Publication: #ANP00100 Printed in Slovenia, 08/2000
We reserve the right to alter specification details etc. without prior notice or liability!
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Courtesy of Dr. Romana Marinsek-Logar, Biotechnical Faculty, Zootechnical Department, University of Ljubljana
For details contact us or visit our home page under Application P001: Partial Purification of a 66 kDa-Endoxylanase and a Broad SpecificityEndoglucanase/Xylanase ...(PDF: 198 KB)
APP_001 BLACK PANTONE 320 CV SYNCOMP
BIA Separations d. o. o.Teslova 30, SI-1000 Ljubljana, Slovenia
Tel.: +386 1 426 5649
Fax: +386 1 426 5650
E-mail: [email protected] Site: www.biaseparations.com
Second separation step shown in Figure B:
Sample: Fraction 4 from the first step (see Figure A);4A, 4B, 4C, 4D and 4E are collected fractions;
Figure C:
Fractions 4A 4D were further analyzed by SDS-PAGE(see Figure C). Purified 66 kDa-Endoxylanase was detectedin fraction 4B.Injection Volume: 7 mLMobile Phase: Buffer A: 20 mM Tris-HCl, pH 7.4;Buffer B: Buffer A + 1 M NaCl, pH 7.4Conditions: Gradient: as shown in Figure B;Flow Rate: 7 mL/min; Detection: UV at 280 nm
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RE
LA
TIV
EA
BSO
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(28
0 nm
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TIME (MINUTES)
% b
uffe
rA
CIM technology is covered by US patents 4889632, 4923610, 4952349 and 5972218. Other patents pending.
19992000 by BIA Separations d. o. o. Publication: #ANP00100 Printed in Slovenia, 08/2000
We reserve the right to alter specification details etc. without prior notice or liability!
beenso easyha
s ne
ver
Scal
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in
chromatography
1999 2000 2001 2002 2003
Figure B:
215 kDa
116 kDa97 kDa
66 kDa
45 kDa
29 kDa
Immunization antibodies
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using CIMtechnology!