Bio-Rad Bio-Gel P Polyacrylamide Gel - Manual (Page 3)

Section 1
Introduction
1.1 Background
The Profinia™ protein purification system provides a complete line of
consumables that interface with an automated, user-friendly instrument for
the purification of polyhistidine- and GST- tagged proteins. The system
has been designed to take standard chromatography buffers, resins,
reagents, and instrumentation and package them into a complete, kit-based,
easy-to-use platform. Four main types of applications currently used in the
Profinia system have kits: native IMAC, denaturing IMAC, GST, and
desalting. The reagents and consumables available for each of these
applications include prepackaged buffers, IMAC and GST affinity media in
prepacked cartridges, and antibody detection tools. Other methods such
as antibody, Profinity eXact™ tag, MBP tag, and Strep-tag purifications,
can also be performed on the Profinia system without kits. Refer to
bulletins 5701, 5725, 5741, and 5744 for buffer formula. See the Ordering
Information section for a complete list of Profinia products.
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 solutions (affinity, desalting*,
cleaning, and storage solutions), and reagents (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 and cartridge
cleaning buffer kit contains 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 1
summarizes the complete line of buffer and purification kits available for
the Profinia system.
* For denaturing IMAC applications, stepwise dialysis is the suggested method for desalting and
buffer exchange and for renaturing proteins.
Important note: Do not use GST lysis and wash buffer for IMAC purification because GST lysis
buffer and wash buffer contain a high concentration (10 mm) of EDTA that will damage the IMAC
cartridge.
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