Bio-rad Affi-Gel Blue Gel User Manual

Affi-Gel®Blue GelInstruction ManualCatalog Numbers153-7301153-7302LIT590B 9/3/98 1:30 PM Page A

Table 2. Affi-Gel Blue Gel Enzyme ApplicationsEnzyme Source Eluant Reference1-aminocyclopropane- Tomato pericarp tissue 50 mM KPO4, 411-carboxylate sy

9nsEluant Referencesue 50 mM KPO4, 415 µM pyridoxal phosphate0.18 M NaCl 30.6 M KCl in 50 mM Tris-HCl 350.4-3.0 M KCl 63 M NaCl, 0.02 M phosphate 371

Table 2. Affi-Gel Blue Gel Enzyme Applications(continued)Enzyme Source Eluant ReferenceGlutamine synthetase Salmonella typhimurium 5 mM ATP 8Glyoxalas

11nsEluant Referenceurium 5 mM ATP 80-0.2 M KCl 19s 1 mM NADPH, 2 mM GMP 210.05-0.75 M NaCl 52 mM NADP 162 mM NADP+ 130.25 M NaCl 7Triton X-100, NaCl,

12Suggested Procedure for Enzyme Purification1. Prepare a column of Affi-Gel blue gel, 100-200 mesh.A 5 ml bed volume for every 20 mg of protein to be

7. Regenerate the column by washing it with 2 bedvolumes of 2 M guanidine HCl or 1.5 M NaSCN,followed by 2 bed volumes of the starting buffer.3.4 Puri

Section 4Storage ConditionsThe Affi-Gel blue gel should be stored at 4 °C in0.02% sodium azide.Section 5References1. Thompson, S. T., et al., Proc. Na

9. Meadows, G. G. and Cantwell, G. S., Res. Comm. inChemical Pathology and Pharmacology, 30, 535 (1980).10. Tomasselli, A. G. and Noda, L. H., Eur. J.

23. Sharma, R. K., et al., J. Biol. Chem., 255, 5916 (1980).24. Wescott, K. R., et al., Proc. Nat. Acad. Sci. USA, 76, 204(1979).25. Wallace, R. W., e

39. Butler-Ransohoff, J. E., et al., Proc. Nat. Acad. Sci. USA,85, 4276 (1988).40. Wold, M. W. and Kely, T., Proc. Nat. Acad. Sci. USA, 85,2523 (1988)

Table of ContentsSection 1 Introduction... 1Section 2 Product Description... 3Sec

Section 7Technical InformationFor additional information and technical assistance,contact your local Bio-Rad representative or callTechnical Service a

Bio-Rad Laboratories, 2000 Alfred Nobel Dr., Hercules, CA 94547LIT-590 Rev BLIT590B 9/3/98 1:30 PM Page BC1

Section 1IntroductionAffi-Gel blue affinity gel is a beaded, crosslinkedagarose gel with covalently attached Cibacron®BlueF3GA dye. It contains ≥1.9 m

Fig. 1. Cibacron blue coupled to agarose.Affi-Gel blue gel is supplied ready to use as anaqueous slurry of fully hydrated gel. It is available intwo c

Section 2Product DescriptionMatrix Bio-Gel A-5m agarose gelParticle Sizes 150-300 µm (50-100 mesh)80-150 µm (100-200 mesh)Shipping Medium 0.05% NaN3Fu

Section 3Instructions for Use3.1 Sample PreparationProper adjustment of the sample pH and ionicstrength is critical for consistent and reproducible re

is so strong that a high concentration of salt or chaotropicreagent is required to desorb the albumin. Other serumproteins either do not bind to Affi-

3. Equilibrate the serum sample in buffer A bydialyzing overnight, or by rapid column desalting ina column of Bio-Gel P-6 DG gel, on an Econo-Pac10DG

biospecifically to the Cibacron Blue F3GA dye.1Inmany cases, the enzyme can be eluted from the Affi-Gelblue gel with a specific nucleotide cofactor. O