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Premium PLUS Expression Kit for MS

Product Name Catalog No. Unit Qty Storage
Premium PLUS Expression Kit for MS EDX-PLUS-MS 1 set 16 reactions -80
This product is shipped on dry ice: Store at -80 upon arrival.
Description

The Premium PLUS Expression Kit for MS provides premixed reagents to perform cell-free protein expression reactions for preparation of [13C, 15N]-L-Lys and [13C, 15N]-L-Arg labeled proteins. Each kit contains the reagents to perform 16 SP6-driven mRNA expression and wheat germ protein labeling reactions.

Intended Use

Preparation of [13C, 15N]-L-Lys and [13C, 15N]-L-Arg labeled proteins for use in protein mass spectrometry experiments. Heavy isotope labeled proteins may be used as controls in sample preparation, for determining digestion efficiency, or as standards for protein quantification.

Key Features:
Labeled proteins are obtained in a fast and convenient three-step process:

Protein Expression made easy: Obtain Template > Express Labeled Protein > Confirm Expression!

Reaction Steps
  1. Transcription using either plasmid DNA template or linear DNA template, incubation at 37 for 1 hour (plasmid DNA) or 4 hours (linear DNA).
  2. Translation of target protein by bilayer reaction, incubation at 15 for 20 hours (plasmid DNA) or 24 hours (linear DNA).

Apparatus required for reaction

Incubator (Transcription at 37 , translation at 15 )

Example Data:

Kit Content:
Reagents Specification Volume
pEU-E01-MCS vector expression vector 5 µL
pEU-E01-DHFR vector positive control 5 µL
PCR primer set Primers SPU and deSP6E01 100 µL ea.
Transcription Premix LM   18 µL x 16
WEPRO9240 for MS1   10 µL x 16
SUB-AMIX SGC2 for MS   206 µL x 16
Aluminum seals For covering reaction cups  

Notes:
(*1) Dedicated wheat germ extract for protein labeling based on the WEPRO7240
(*2) contained in single-break strip wells consisting of 8 wells, which are also used as reaction cups

Shelf-life: 1 year
This product is produced in collaboration with Cambridge Isotope Laboratories, Inc.

Support
Instruction Manual
Selected References

FLEXIQuant: A novel tool for the absolute quantification of proteins, and the simultaneous identification and quantification of potentially modified peptides.
Singh, S., Springer, M., Steen, J., Kirschner, M.W., and Steen, H. (2009)
J Proteome Res., 8, 2201 - 2210
FLEXIQinase, a mass spectrometry-based assay, to unveil multikinase mechanisms.
Singh SA, Winter D, Bilimoria PM, Bonni A, Steen H, and Steen JA. (2012)
Nat Methods. Apr 8;9(5):504-8.
A practical guide to the FLEXIQuant method.
Singh S, Kirchner M, Steen JA, Steen H. (2012)
Methods Mol Biol. 2012;893:295-319

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