2012 Promega catalogue

Cell Signaling
HaloTag ® Mammalian Pull-Down Systems
Product Size Cat.# Price ($)
HaloTag ® Mammalian Pull-Down and
Labeling System 24 reactions G6500 521.00
HaloTag ® Mammalian Pull-Down System 24 reactions G6504 284.00
HaloTag ® Control Vector 20 µg G6591 231.00
Available Separately
Protease Inhibitor Cocktail, 50X 1 ml G6521 87.00
Mammalian Lysis Buffer 40 ml G9381 58.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The HaloTag ® Mammalian Pull-Down and Labeling
System (Cat.# G6500 and G6504) is designed to capture and purify intracellular
binary and higher order protein complexes, including transient or weakly
interacting partners. This system includes the HaloTag ® TMRDirect Ligand,
which allows correlative cellular localization and real-time imaging studies with
the same genetic construct as a means to further understand overall protein
function. Cat.# G6504 does not include the TMRDirect Ligand.
The HaloTag ® Control Vector provides protein expression of the HaloTag ®
protein in mammalian cells, E. coli or in vitro expression systems dependent
on human cytomegalovirus (CMV) intermediate early enhancer, T7 or SP6
RNA polymerase promoters. It can be used as a control for any HaloTag ®
experimental system and can be used for both stable and transient HaloTag ®
expression in mammalian cells; for stable expression, co-transfection with a
vector containing a selectable marker is required.
The Protease Inhibitor Cocktail, 50X, is a mixture of six different protease
inhibitors with different target protease specificities. This product is provided in
a freeze-dried format and can be reconstituted using either 100% ethanol or
DMSO.
POI
HT
HaloTag ®
fusion construct POI
HT
Protein of Interest
HaloTag ® Protein
Transfection
or stable
expression
TEV
cleavage
POI
For complete and up-to-date product information visit: www.promega.com/catalog
POI
POI
POI
HT
HT
HT
HaloLink
Resin
SDS
elution
The Mammalian Lysis Buffer is designed for use with HaloTag ® Mammalianbased
expression systems such as the HaloTag ® Mammalian Pull-Down and
Labeling Systems (referenced here) as well as the HaloCHIP System (Cat.#
G9410). Formulation consists of 50mM Tris-HCl, 150mM NaCl, 1% Triton ®
X-100 and 0.1% sodium deoxycholate (pH 7.5).
Features:
• Rapid, Efficient and Covalent Capture of Binary and Higher Order
Complexes Directly from Lysates: Improved capture of protein partners,
including transient interactions.
• High Purity and Low Background: Improved accuracy in identification
of proteins; covalent attachment allows bait protein to remain behind if
desired.
• Ability to Fluorescently Label the Same Genetic Fusion: Correlate
complex capture with cellular localization.
• Compatibility with All Downstream Methods of Analysis: Freedom to
identify complexes in variety of applications including mass spectrometry.
Storage Conditions: Store the 10X TBS Buffer and SDS Elution Buffer at
room temperature. Store the HaloLink Resin and Mammalian Lysis Buffer at
4°C. Store the Protease Inhibitor Cocktail and HaloTag ® TMRDirect Ligand
at –20°C.
Protocol Part#
HaloTag ® Mammalian Pull-Down and Labeling Systems Technical
Manual
TM342
Express HaloTag ® fusion protein
and assemble protein complex.
Lyse cells.
Capture protein complex on
HaloLink Resin.
Wash HaloLink Resin.
Elute with SDS buffer or
cleave with TEV (optional).
Analyze with downstream
application.
Representation of the HaloTag ® mammalian pull-down assay using HaloTag ® fusion protein as bait.
For Expression-Validated HaloTag ® ORF Clones
visit Find My Gene at: www.promega.com/FMG
9033MA
245
14
Protein Purification and Interactions
Section
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