2012 Promega catalogue

Cell Signaling
pGEM ® β-Gal Control DNA
Product Size Cat.# Price ($)
pGEM ® β-Gal Control DNA 20 µg L4731 193.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: pGEM ® β-Gal Control DNA contains the coding sequence of
β-galactosidase downstream of an E. coli wildtype lacZ promoter. pGEM ®
β-Gal Control DNA can be used as a positive control in the E. coli S30 Extract
System for Circular DNA. The wildtype lacZ promoter is not efficient for initiating
transcription from a linear DNA template. Supplied as a 0.5mg/ml solution in
TE buffer.
Storage Conditions: Store at –20°C.
Cell-Based Protein Expression
Regulated Mammalian Expression System
Product Size Cat.# Price ($)
Regulated Mammalian Expression System 1 system C9470 1208.00
Coumermycin A1 5 mg C9451 125.00
Novobiocin Sodium Salt
Available Separately
1 g C9461 47.00
pReg neo Vector 20 µg C9421 386.00
pF12A RM Flexi ® Vector 20 µg C9431 494.00
pF12K RM Flexi ® Vector 20 µg C9441 494.00
C9421, C9470, C9431, C9451, C9441 For Research Use Only. Not for Use in Diagnostic
Procedures. C9461 For Research Use Only. Not for Use in Therapeutic or Diagnostic
Procedures.
Description: The Regulated Mammalian Expression System features low
basal levels, robust and rapid induction, and downregulation of gene expression
in mammalian cells. The Regulated Mammalian Expression System is based
on a novel on/off switch that relies on the rapid and sensitive modulation by
coumerin-related compounds of a chimeric transactivator protein. Nanomolar
concentrations of the antibiotic coumermycin promote homodimerization of a
chimeric transactivator that, in turn, binds to lambda operator sequences located
upstream of a minimal promoter driving transcription of coding sequences for a
protein of interest. The levels of protein expression can be regulated by adjusting
the coumermycin concentration. More significantly, this expression can be
promptly and effectively switched off by adding novobiocin, which acts as an
antagonist by dissociating the dimerized transactivator protein.
The protein coding region of interest is cloned into either the pF12A RM Flexi ®
Vector or pF12K RM Flexi ® Vector, both of which are specially designed for
Regulated Mammalian (RM) protein expression. These vectors incorporate
regulatory promoter sequences upstream of the protein-coding region and are
compatible with the Flexi ® Vector System. In transient transfection paradigms,
the pF12A or pF12K RM Flexi ® Vector containing the protein-coding region
of interest is co-transfected into mammalian cells together with the pReg neo
Vector. The pReg neo Vector is designed to express a chimeric transactivator
protein that interacts with the regulatory promoter region in the pF12A and
pF12K RM Flexi ® Vectors in a regulated fashion in response to coumermycin
and novobiocin. Additionally, the pReg neo Vector encodes a neomycin
phosphotransferase gene that allows stable cell selection and generation with
the antibiotic G-418.
Features:
• Enhanced Data: High level of controlled induction combined with low
basal protein expression.
• Regulated Expression: Dose-response induction of protein expression;
rapid and sensitive on/off switch for protein expression.
• Versatility: Compatible with other Flexi ® Vectors.
Storage Conditions: Store at –20°C.
Protocol Part#
Regulated Mammalian Expression System Technical Manual TM289
For complete and up-to-date product information visit: www.promega.com/catalog
Single Step (KRX) Competent Cells
Product Size Cat.# Price ($)
Single Step (KRX) Competent Cells 20 × 50 µl L3002 330.00
L-Rhamnose Monohydrate 10 g L5701 70.00
50 g L5702 272.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The Single Step (KRX) Competent Cells are designed for efficient
transformation and tightly controlled protein expression. This one strain
consolidates the best attributes of these two steps into one strain to evaluate
protein expression in E. coli.
Transformation efficiencies are greater than 108 cfu/μg, similar to other highly
competent cells. The single step cells are available in single transformation size
(50μl). KRX also can be used for blue/white screening.
Single Step (KRX) is an E. coli K strain that contains a chromosomal copy of
the T7 RNA polymerase driven by a rhamnose promoter (rhaBAD) to provide
dramatic control of the proteins expressed via a T7 promoter. Pre-induced
expression protein levels are significantly lower than those of BL21(DE3)derived
strains. This feature facilitates cloning and expression of proteins toxic
to E. coli.
Genotype: [F´, traD36, ∆ompP, proA + B + , lac Iq , ∆(lacZ)M15] ∆ompT, endA1,
recA1, gyrA96 (Nalr ), thi-1, hsdR17 (r –
k , mk
+ ), e14 – (McrA – ), relA1, supE44,
∆(lac-proAB), ∆(rhaBAD)::T7 RNA polymerase.
Features:
• Save Time: In two days, you can transform your vector into the Single
Step (KRX) Competent Cells and be ready for protein expression.
• Controlled Protein Expression: For overall expression of cloned
proteins, the Single Step (KRX) Competent Cells provide dramatic control of
expressed protein-coding regions.
• Achieve High Yields: Protein expression levels were shown to be as high
as or higher than levels expressed in BL21(DE3)-derived strains.
Protocol Part#
Single Step (KRX) Competent Cells Technical Bulletin TB352
229
13
Protein Expression and Analysis
Section
Contents
Table of
Contents