Purification kits – Bio-Rad Profinia™ Protein Purification Instrument User Manual

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Section 10
Profinia

Purification Kits

10.1 Introduction

10.1.1 Background

The Profinia protein purification system includes a complete line of consumables that
interface with the instrument for the purification of affinity-tagged proteins. The consumables
consist of standard chromatography buffers, resins, reagents, and accessories that have
been packaged into a complete, kit-based, and easy-to-use platform. Reagents and
purification kits have been developed for the four main types of system applications —
native IMAC, denaturing IMAC, GST, and desalting — and include bacterial lysis reagents,
prepackaged buffers, IMAC and GST affinity media in prepacked cartridges, and antibody
detection tools. See Appendix B (Ordering Information) for a complete list of Profinia
products.

10.1.2 Product Information

Two types of kits can be used for the purification of affinity-tagged proteins: Profinia
purification kits and Profinia buffer kits. The all-inclusive Profinia purification kits include two
1 ml affinity and two 10 ml desalting cartridges, in addition to a complete set of buffers and
solutions (affinity, desalting*, cleaning, and storage solutions) and reagents (urea and
glutathione) required for affinity and desalting applications. Profinia buffer kits are an
appropriate choice for those users requiring additional buffers and cleaning reagents, but
are not supplied with the affinity or desalting cartridges. The desalting buffer and separations
kits contain solutions and cartridges specific for desalting or buffer exchange.

Each kit contains sufficient reagents to perform 10 ml of affinity purifications, followed by
100 ml of desalting separations. This can be broken up into ten individual 1 ml affinity
cartridge runs, two 5 ml affinity cartridge runs, or a single 5 ml and five 1 ml cartridge runs.
The cartridges for affinity/desalting applications are provided as 1 ml/10 ml pairs. Larger
5 ml/50 ml affinity/desalting cartridges can be purchased separately for larger-scale
purification runs. Because each ml of IMAC or GST resin typically yields ~10 mg of purified
protein, each buffer or purification kit typically yields ~100 mg of purified protein. Table 10.1
summarizes the complete line of purification and buffer kits available for the Profinia system.

* Denaturing IMAC kits do not contain desalting cartridges or desalting buffer; stepwise dialysis is the suggested
method for desalting and buffer exchange and for renaturing proteins purified under denaturing conditions.

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