2012 Promega catalogue

Life
Science
Catalog
2012
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Cell Signaling
252
HisLink Spin Protein Purification System
Product Size Cat.# Price ($)
HisLink Spin Protein Purification System 25 reactions V1320 335.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The HisLink Spin Protein Purification System provides a
simple, quick and robust method of purifying polyhistidine- or HQ-tagged
expressed proteins from E. coli using either a centrifuge- or vacuum-based
method.
Proteins can be purified directly from culture medium containing bacterial cells
expressing polyhistidine- or HQ-tagged protein. The bacterial cells are lysed
using the FastBreak Cell Lysis Reagent, followed immediately by the addition
of the HisLink Protein Purification Resin to the culture. The addition of these
reagents results in simultaneous bacterial lysis and binding of the polyhistidine-
or HQ-tagged proteins. Samples are then transferred to a Spin Column, where
unbound protein is washed away and the target protein of interest is recovered
by elution.
Features:
• Versatile: Choose between vacuum or centrifugation formats.
• Convenient: Complete system containing all necessary components.
• Productive: High binding capacity per column.
Storage Conditions: Store the system at 4°C. Store Spin Columns,
Collection Tubes and FastBreak Cell Lysis Reagent at room temperature.
After reconstitution, store the DNase I in aliquots at –20°C.
Protocol Part#
HisLink Spin Protein Purification System Technical Manual TM281
Biotin-Avidin Protein Purification
SoftLink Soft Release Avidin Resin
Product Size Cat.# Price ($)
SoftLink Soft Release Avidin Resin 1 ml V2011 250.00
For Research Use Only. Not for Use in Diagnostic Procedures.
5 ml V2012 1004.00
Description: SoftLink Avidin Resin can be used for the isolation and
purification of biotinylated molecules. SoftLink Resin is a rigid, methacrylate
polymeric gel filtration matrix, functionalized with covalently bound, monomeric
avidin. Monomeric avidin binds biotin with a K d value of 10 –7 M, allowing
reversible binding of bound biotinylated proteins under mild elution conditions.
Native, or tetrameric, avidin binds biotin with a very strong affinity (K d = 10 –15 M),
which in turn requires strong denaturing conditions for eluting bound material.
Monomeric avidin allows the specificity of capture but also the mildness of
release appropriate for the purification of sensitive biological materials.
Features:
• Sensitive: Binds 20–40nmol of biotinylated protein per milliliter of resin.
• Easy to Use: Bound biotinylated molecules can be eluted under mild
nondenaturing conditions (5mM biotin).
• Versatile: Retains biotin binding ability following exposure to a wide range
of pH, low or high ionic strength, 6M guanidine and 1% SDS.
• Reusable: Regenerates at least 10 times without loss of binding capacity.
• Robust: Supports high flow rates (300cm/hour) and centrifugal forces
(1,500 × g) in batch applications.
• Flexible: Purifications by batch or column method.
Storage Conditions: Store at 4°C.
Protocol Part#
SoftLink Soft Release Avidin Resin Product Information 9PIV201
PinPoint Xa Protein Purification System
Product Size Cat.# Price ($)
PinPoint Xa Protein Purification System
Available Separately
1 system V2020 1130.00
PinPoint Xa-1 Vector 10 µg V2031 303.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The PinPoint Xa Protein Purification System is designed for
the production and purification of fusion proteins that are biotinylated in vivo.
The DNA coding for the protein of interest is cloned into a PinPoint Vector
downstream of a sequence encoding a peptide that becomes biotinylated in
vivo. Biotinylated fusion proteins are produced in E. coli and are affinity-purified
using the SoftLink Soft Release Avidin Resin. This proprietary resin allows
elution of the fusion protein under nondenaturing conditions. The PinPoint
Vectors feature the encoded endoproteinase Factor Xa (pronounced “ten a”)
proteolytic site that provides a way to separate the purification tag from the
native protein, and the vectors carry a convenient multiple cloning region for
ease in construction of fusion proteins.
The system contains vectors in all possible sense reading frames, an avidinconjugated
resin, Streptavidin-Alkaline Phosphatase, a purification column
and biotin. The PinPoint Xa Control Vector contains the chloramphenicol
acetyltransferase (CAT) gene and is provided as a means of monitoring protein
expression, purification and processing conditions. The system generally yields
1–5mg of protein per liter of culture.
Features:
• In vivo Biotinylation Tag: Allows purification of fusion proteins; many
proteins produced have been soluble.
• Easy to Use: Purification of biotinylated proteins with the SoftLink Resin
can be performed by column or batch purification.
• Easy Detection: Streptavidin Alkaline Phosphatase can be used to detect
the biotinylated fusion protein in a pseudo-Western format to monitor
purification.
• Flexible: PinPoint Vectors are supplied for all reading frames.
• Gentle Release Conditions: SoftLink Resin allows release of the
fusion protein under nondenaturing conditions.
• tac Promoter: Allows tightly regulated expression.
Storage Conditions: Store the PinPoint Purification Column at room
temperature. Store all remaining components at 4°C. The vectors may be
stored at –20°C.
Protocol Part#
PinPoint Xa Protein Purification System Technical Manual TM028
PinPoint Vector Sequencing Primer
Product Size Cat.# Price ($)
PinPoint Vector Sequencing Primer
For Research Use Only. Not for Use in Diagnostic Procedures.
2 µg V4211 131.00
Description: The PinPoint Vector Sequencing Primer is designed for
sequencing inserts cloned into the PinPoint Xa Vectors (components of
Cat.# V2020). The primer hybridizes upstream of the Factor Xa site at
nucleotides 325–343, approximately 40–50 base pairs upstream of
the multiple cloning region and can be used to determine if an insert is
cloned in-frame with the biotinylation purification tag of the PinPoint
Xa Vectors. The sequence of the PinPoint Vector Sequencing Primer is
5´-d(CGTGACGCGGTGCAGGGCG)-3´. It is supplied dried.
Features:
• Performance Tested: The PinPoint Vector Sequencing Primer is tested
in double-stranded sequencing reactions with circular PinPoint Vectors.
Storage Conditions: Store at –20°C.
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