ReadyPrep™ Reduction-Alkylation Kit

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Overview

The ReadyPrep reduction-alkylation kit reduces and irreversibly blocks the formation of inter- and intramolecular disulfide bonds prior to isoelectric focusing (IEF).
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Description

Description

The ReadyPrep reduction-alkylation kit reduces and irreversibly blocks the formation of inter- and intramolecular disulfide bonds prior to isoelectric focusing (IEF). Disulfide bond formation is particularly problematic for basic proteins because of the increased rate of their formation in an alkaline environment. Moreover, many reducing agents become negatively charged during IEF and migrate off the IPG strip, thus allowing disulfide bonds to re-form (Görg 2000, Hoving 2002).

The ReadyPrep reduction-alkylation kit:

  • Eliminates the potential for disulfide bond formation during IEF
  • Reduces disulfide bonds with tributylphosphine (TBP) and alkylates the sulfhydryl groups with iodoacetamide (Herbert et al. 2001)
  • Provides reagents for treatment of 100 samples (300 µl each, containing up to 10 mg/ml of protein)
  • Performs treatment directly in the rehydration/sample buffer, in less than 2 hours, and with very little hands-on time
  • Is compatible with other ReadyPrep sample preparation kits and buffers

References
Görg A et al. (2000). The current state of two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis 21, 1,037–1,053.
Herbert B et al. (2001). Reduction and alkylation of proteins in preparation of two-dimensional map analysis: Why, when and how? Electrophoresis 22, 2,046–2,057.
Hoving S et al. (2002). Preparative two-dimensional gel electrophoresis at alkaline pH using narrow range immobilized pH gradients. Proteomics 2, 127.

Ordering

Ordering

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1632090
100 preps, includes 3 x 0.6 ml 200 mM TBP, 3 empty glass vials for TBP storage, 5 x 56 mg iodoacetamide, 2 ml alkylation buffer, instruction manual; for 100 x 300 µl samples containing up to 10 mg/ml protein

Kits & Reagents

Kits & Reagents

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1632091
500 ml, ultrapure water for use with ReadyPrep fractionation and general purpose cleanup kits
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(Discontinued) Iodoacetamide
1632109
30 g, alkylating agent for use in protein sample preparation applications
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1632101
0.6 ml, 200 mM, reducing agent for use in protein sample preparation applications

Documents

Documents

Number Description Options
4110063
Instruction Manual, ReadyPrep™ Reduction-Alkylation Kit, Rev A