2013 Promega catalogue

Cell Signaling
HaloTag ® Protein Purification System
Product Size Cat.# Price ($)
HaloTag ® Protein Purification System 1 each G6280 926
HaloTag ® Protein Purification System Sample Pack 1 each G6270 313
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The HaloTag ® Protein Purification System is designed to purify
proteins fused to HaloTag ® , a novel protein tag that enhances the expression
and solubility of recombinant proteins. HaloTag ® Technology enables the
covalent, efficient and specific capture of a protein of interest onto HaloLink
Resin, thus overcoming the equilibrium-based limitations associated with affinity
tags (i.e., poor capture of proteins expressed at low levels and protein loss
during washing of the purification resin.
The HaloTag ® technology offers a quick and convenient way to test protein
expression of HaloTag ® fusion proteins as well as monitor the efficiency of
immobilization to HaloLink Resin by labeling with fluorescent HaloTag ® TMR
Ligand followed by SDS-PAGE analysis. Guidelines can be found in the Halo-
Link Resin Technical Manual #TM250, the HaloLink Protein Array Technical
Manual #TM310, and the HaloCHIP System Technical Manual #TM075.
Outline of Procedure
The HaloTag ® protein, a 34kDa mutated hydrolase, covalently attaches to
HaloLink Resin via an immobilized chloroalkane ligand. TEV Protease cleaves
the target protein from the HaloLink Resin. The TEV Protease, which has an
N-terminal (HQ) tag, is removed from the protein of interest using HisLink
Resin, and the purified protein of interest is recovered. The appropriate vector
that encodes the HaloTag ® protein and expresses protein optimally in E. coli
is pFN18A HaloTag ® T7 Flexi ® Vector (G2751) or pFN18K HaloTag ® T7 Flexi ®
Vector (G2681). These vectors can be purchased separately.
Features:
• Experience Superior Yield, Purity and Specific Activity of Soluble,
Functional Proteins Compared to His-Tag, GST and MBP Affinity
Tags: Specific and covalent HaloTag ® fusion protein capture and immobilization
on HaloLink Resin.
• Achieve Enhanced Target Protein Expression in Prokaryotic, Mammalian
and Cell-Free Systems: Proteins are expressed as HaloTag ®
fusion proteins.
• Purify Poorly Expressed Fusion Proteins: Rapid, specific and covalent
capture of HaloTag ® protein onto HaloLink Resin is a nonequilibrium
process.
• Efficiently Recover Tag-Free Target Protein using TEV Protease
Cleavage: Optimized TEV protease recognition site within the interconnecting
polypeptide separating the HaloTag ® protein and the fusion
partner. HaloTag ® protein remains immobilized on the resin due to covalent
capture.
• Save Time: One buffer compatible with downstream applications for all
purification steps.
• Perform Easy In-Gel Detection and Quantification of Protein
Expression Levels with Fluorescent HaloTag ® Ligands: Highly stable
HaloTag ® protein-ligand interaction permits boiling with SDS sample buffer
followed by resolving on SDS-PAGE.
Storage Conditions: Store the HaloLink Resin and HisLink Resin at 4°C.
Do not freeze the resins. Store the TEV Protease at –20°C.
Protocol
HaloTag ® Protein Purification System Technical Manual
Part#
TM312
8
Epigenetics
Schematic of the HaloTag ® Mammalian Protein Purification System protocol. Streamlined purification process leads to higher purity and recovery of
recombinant proteins from cultured cells.
Section
Contents
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