Bio-rad Affi-Gel Hz Hydrazide Gel User Manual

Affi-Gel®HzImmunoaffinityKitInstructionManualCatalog Number153-6060For Technical ServiceCall Your Local Bio-Rad Office orin the U.S. Call 1-800-4BIORA

4. Transfer the gel buffer slurry to a coupling reaction tube. If lessthan 5 ml of gel is to be used for coupling, wash only the vol-ume of gel to be

= (mg IgG/ml) x dilution factor x sample volume = total IgG[total protein before coupling] - [total uncoupled protein (eluant + 0.5 M NaClwash)]= [tot

5.2 Sample Application1. Sample is applied to the immobilized IgG column. Samplesshould be free of particulates. Complex samples should be dilut-ed i

Chaotropic Elution4 M NaSCN6 M urea5 M guanidine-HC1Elution StrategiesThe preceding eluants generally do not denature antibodies.*Eluant must be compa

CatalogNumber Product Description130-0150 Bio-Gel HT Hydroxyapatite, 250 ml156-0030 Macro-Prep High S Support, 100ml156-0090 Macro-Prep t-Butyl HIC Su

Bio-Rad LaboratoriesU.S. (800) 4BIORAD California (510) 741-1000 Australia 02-9914-2800 Austria (1) 877 89 01 Belgium 09-385 55 11 Canada (905) 712-2

Table of ContentsSection 1 Introduction...11.1 Immunoaffinity Coupling and Usefulness of

Section 1IntroductionThe Affi-Gel Hz immunoaffinity kit is a unique approach toIgG coupling to an agarose support matrix for affinity purification.Thi

1.2 Hydrazide Coupling Chemistry1.3 Diagram of Kit ComponentsMaterials and Equipment Not Included in this KitEssential15 ml screw cap test tubes - p

OptionalBio-Rad Protein Assay Kit I (catalog number 500-0001)Section 2Monoclonal AntibodiesDue to the unique specificity of monoclonal antibodies, cou

matography may also be used to remove most serum componentsexcept transferrin.IgG in ascites fluid and serum may be purified by Affi-Gel pro-tein A ag

in affinity chromatography. 4.1A Dilution of Affi-Gel Hz 10x Coupling Buffer1. Dilute Affi-Gel Hz 10x coupling buffer 1:10 with distilled,deionized w

6. The Econo-Pac 10DG column should be washed with at least 20ml of diluted coupling buffer, pH 5.5, to regenerate for desalt-ing use after oxidation.

6. Immediately proceed to desalting, Section 4.3.4.3 Desalting ProcedureImmediately after the 1 hour oxidation, it is necessary to removethe sodium p