2012 Promega catalogue

Life
Science
Catalog
2012
Worldwide Contact List
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Contents
Table of
Contents
Cell Signaling
230
BL21(DE3)pLysS Competent Cells
Product Size Cat.# Price ($)
BL21(DE3)pLysS Competent Cells, >10 6 cfu/µg 1 ml L1191 238.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: BL21(DE3)pLysS Competent Cells allow high-efficiency protein
expression of any gene that is under the control of a T7 promoter and has a
ribosome binding site. BL21(DE3)pLysS is lysogenic for λ-DE3, which contains
the T7 bacteriophage gene I, encoding T7 RNA polymerase under the control of
the lac UV5 promoter. BL21(DE3)pLysS also contains a plasmid, pLysS, which
carries the gene encoding T7 lysozyme. T7 lysozyme lowers the background
expression level of target genes under the control of the T7 promoter but does
not interfere with the level of expression achieved following induction by IPTG.
One milliliter is sufficient for ten transformations.
Genotype: F–, ompT, hsdS B (r B–, m B–), dcm, gal, λ(DE3), pLysS, Cm r .
Features:
• T7 Promoter Expression: Contains an IPTG-inducible gene for T7 RNA
polymerase.
• Convenient: A time-saving alternative to making competent cells.
Storage Conditions: Store at –70°C.
Protocol Part#
E. coli Competent Cells Technical Bulletin TB095
A.
Normalized luminescence (RLU/OD)
B. 10,000
Induction Ratio
10,000
9,000
8,000
7,000
6,000
5,000
4,000
3,000
2,000
1,000
0
1,000
100
10
1
Normalized luminescence (RLU/OD)
BL21(DE3) BL21(DE3)
pLysS
14
140
120
100
80
60
40
20
0
8
BL21(DE3) BL21(DE3)
pLysS
Pre-Induction Expression
Roset t a 2
pLysS
BL21(DE3)
pLysS
BL21(DE3)
43
Rosetta 2
pLysS
KRX
Rosetta 2
pLysS
1,700
Pre-induction and post-induction expression levels of firefly
luciferase. Cells were transformed with the pF1K T7 Flexi ® Vector
containing the firefly luciferase gene. Cultures were grown at 37°C to
an optical density (O.D. 600) of 0.8–1.0 and then moved to a 25°C incubator
shaker. When cultures reached an O.D. 600 of 1.0–1.5, protein expression
was induced using either 0.1% rhamnose or 1mM IPTG and grown overnight
at 25°C. Samples for luciferase assays were removed prior to and after
induction. Panel A. Firefly luciferase expression level was determined using
the Bright-Glo Luciferase Assay Reagent. Pre- and post-induction firefly
luciferase expression levels were normalized to cell number (n = 3). Panel B.
Induction ratios were calculated by dividing the post-induction luminescence
values by the pre-induction values.
KRX
KRX
6032MA
Mass Spectrometry Analysis
ProTEV Plus
Product Size Conc. Cat.# Price ($)
ProTEV Plus 1,000 u 5 u/µl V6101 165.00
8,000 u 5 u/µl V6102 1163.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: ProTEV Plus is an engineered form of TEV protease, a highly
specific proteolytic enzyme that cleaves within a seven-amino-acid sequence.
It can be used to cleave protein fusions that have been engineered to contain
the seven-amino-acid sequence at the desired cleavage site. ProTEV Plus also
contains an HQ tag located at the N-terminus of the protein, which allows it to
be immobilized on affinity resins and removed from the cleavage reaction.
Features:
• Active Over a Wide Range of pH and Temperatures: Cleave individual
fusion proteins using optimal conditions to maintain activity and correct
conformation.
• HQ-Tagged: Convenient removal of ProTEV Plus using Ni-based affinity
resins after cleavage.
• Cleaves Fusion Proteins Directly in Solution or Immobilized on Affinity
Resins: ProTEV Plus is easy to use in multiple experimental formats.
Storage Conditions: Store at –20°C.
Protocol Part#
ProTEV Plus Protease Product Information 9PIV610
ProteaseMAX Surfactant, Trypsin Enhancer
Product Size Cat.# Price ($)
ProteaseMAX Surfactant, Trypsin Enhancer 1 mg V2071 100.00
5 × 1 mg V2072 395.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: ProteaseMAX Surfactant, Trypsin Enhancer, is designed to improve
in-gel and in-solution protein digestion. ProteaseMAX Surfactant ensures
fast and efficient protein digestion with proteases such as Trypsin, Chymotrypsin
and Lys-C. For in-gel protein digestion, ProteaseMAX Surfactant offers time
and labor savings. Digestion step is complete in 1 hour, and the surfactant
provides concurrent extraction of peptides from gels, eliminating the need for
post-digestion peptide extraction. The surfactant also improves recovery of longer
peptides that are retained in the gel under a standard extraction protocol.
For in-solution digestions, ProteaseMAX Surfactant solubilizes proteins,
including difficult proteins (i.e., membrane proteins), and enhances protein
digestion by providing a denaturing environment prior to protease addition.
ProteaseMAX Surfactant degrades over the course of a digestion reaction,
yielding products that are compatible with downstream methods such as mass
spectrometry (MS) and liquid chromatography (LC). ProteaseMAX Surfactant
can be used with existing in-gel or in-solution digestion protocols.
Features:
• No Requirement for Peptide Extraction following In-Gel Digestions:
Increase the number of samples processed.
• One-Hour Digestion: Save time by avoiding overnight digestions.
• Improved Peptide Recovery from Gels: Increase in protein sequence
coverage, thus increasing confidence of protein identification.
• Enhanced Protein Solubilization: Solubilize complex proteins such as
membrane proteins at room temperature, avoiding high temperatures and
preventing precipitation.
• Self-Degradable: Use directly for mass spectrometry analysis without
additional inactivation steps such as heating or acid treatment.
Storage Conditions: Store lyophilized ProteaseMAX Surfactant at –20°C.
Protocol Part#
ProteaseMAX Surfactant, Trypsin Enhancer Technical Bulletin TB373
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