Waters Protein-Pak Epoxy-Activated Affinity Products Manuel d'utilisateur télécharger pdf

[ Care and Use ManUal ]

Protein-Pak Epoxy-Activated Affinity 1

Contents

I. IntroduCtIon

a. Chemistry

b. Theory of Affinity Chromatography

c. Description of the Products

II. use of epoxy-ACtIvAted AffInIty pACkIngs

a. Coupling Buffers

b. Coupling Conditions

c. Blocking Procedure

d. Washing Procedure

e. Storage

III. usIng the MICro-ColuMn

a. Preparing the Micro-Column

b. Isolation of the Selected Product

proteIn-pAk epoxy-ACtIvAted AffInIty produCts

I. IntroduCtIon

Waters Protein-Pak™ Epoxy-Activated Affinity products are silica-

based materials with nominal 40 μm particle diameter and nominal

500 Å pore size. The packing is epoxy-activated for ligand coupling.

It is capable of rapid purification of large sample volumes at high

flow rates.

a. Chemistry

Waters Protein-Pak Epoxy Activated Affinity packing is prepared

by bonding an epoxide functionality to silica. The resulting epoxy

groups covalently bond amino, sulfhydryl, and hydroxyl moieties

on small and large ligand molecules including proteins and

pept ides. T he or d er of reac t ivi ty of the l i gan d n u cleop h iles is :

–

> RO

–

> RSH > R

NH > ROH.

b. Theory of Affinity Chromatography

Affinity chromatography is the only chromatographic method in which

specific molecules are isolated on the basis of their bioselective

interaction with an immobilized ligand. Other chromatographic

methods rely on reversible adsorption due to physical and/or

chemical properties of the biomolecule and the support.

Epoxy-activated affinity chromatography can be performed

through the following six steps, as shown in Figure 1. Refer to

Section IV, b Operation, to perform an affinity chromatography

separation as illustrated.

Step 1: Ligand coupling or immobilization

Step 2: Loading sample mixture

Step 3: Adsorbing target material

Step 4: Washing out impurities

Step 5: Eluting target material

Step 6: Re-equilibrating the packing material

1 2 3 4 5 6

[ Care and Use ManUal ] 1

Résumé du contenu

Page 1 - [ Care and Use ManUal ]

[ Care and Use ManUal ]Protein-Pak Epoxy-Activated Affinity 1ContentsI. IntroduCtIona. Chemistryb. Theory of Affinity Chromatographyc. Description of

Page 2

[ Care and Use ManUal ]Protein-Pak Epoxy-Activated Affinity 2Figure 1: Epoxy-Activated Affinity Processc. Description of the ProductsEpoxy-activated p

Page 3

[ Care and Use ManUal ]Protein-Pak Epoxy-Activated Affinity 3Table 1: Guidelines for Coupling to an Epoxy-Activated Packing Material* For insoluble or

Page 4

[ Care and Use ManUal ]Protein-Pak Epoxy-Activated Affinity 4The coupling supernatant and sodium chloride washes may be saved to quantitate the unboun

Page 5

[ Care and Use ManUal ]Protein-Pak Epoxy-Activated Affinity 5c. Storage – 0.01 to 0.1 M sodium phosphate (pH 7.4) – 0.01 to 0.5 M sodium chloride –

Page 6

[ Care and Use ManUal ]Protein-Pak Epoxy-Activated Affinity 6Waters Corporation 34 Maple Street Milford, MA 01757 U.S.A. T: 1 508 478 2000 F: 1 508 87

Commentaires sur ces manuels

Pas de commentaire

Waters Protein-Pak Epoxy-Activated Affinity Products Manuel d'utilisateur

Naviguer en ligne ou télécharger Manuel d'utilisateur pour Équipement pour l'eau Waters Protein-Pak Epoxy-Activated Affinity Products. Waters Protein-Pak Epoxy-Activated Affinity Products User Manual Manuel d'utilisatio

Résumé du contenu

Commentaires sur ces manuels

Produits connexes et manuels pour Équipement pour l'eau Waters Protein-Pak Epoxy-Activated Affinity Products