Bio-rad Profinity eXact™ Purification and Tag Cleavage Con Manual de usuario descargar pdf (Pagina 16)

Problem Possible Cause Solution

Precipation during Binding capacity of Load less sample

purification cartridge exceeded

Protein aggregation Include detergent or

glycerol in buffers

Section 11

Troubleshooting Guide

Problem Possible Cause Solution

Cartridge clogging Particulates in samples Filter all samples and buffers

or slow flow rate or buffers through 0.45 µM filter prior

to application

Sample too viscous Add nuclease to lysate to

degrade DNA

No target protein Low level of target Check expression level by

in eluant protein in starting SDS-PAGE

material

Target protein not Used resin must be

binding regenerated before reuse.

Follow regeneration protocol

in Section 10

Inaccessible tag Clone into pPAL’s Spe I site

Target protein in Intrinsic cleavage Ensure no chloride ions are

flowthrough or wash present in lysate or bind/wash

fractions buffers. Perform purification

at 4°C. Clone into pPAL’s

Spe I site

Uncomplete elution No or slow cleavage Lengthen cleavage incubation

step. Clone into pPAL’s Spe I

site

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Bio-rad Profinity eXact™ Purification and Tag Cleavage Con Manual de usuario Pagina 16