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A 2D card for the Amersham WB system

IP.com Disclosure Number: IPCOM000248647D
Publication Date: 2016-Dec-22
Document File: 2 page(s) / 214K

Publishing Venue

The IP.com Prior Art Database

Abstract

In two-dimensional gel electrophoresis, the first step is to separate proteins according to their isoelectric point, preferably in a gel strip with an immobilized pH gradient, e.g. an IPG DryStrip. Next step is to place this gel strip onto a gel for size separation of the focused proteins, i.e. SDS-PAGE.

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Title: A 2D card for the Amersham WB system

Introduction

In two-dimensional gel electrophoresis, the first step is to separate proteins according to their isoelectric point, preferably in a gel strip with an immobilized pH gradient, e.g. an IPG DryStrip. Next step is to place this gel strip onto a gel for size separation of the focused proteins, i.e. SDS-PAGE.

In the Amersham WB system (AWB), gels for SDS-PAGE are cast between a foil package and a hard plastic part, also having a number of sample wells. The gel is attached to the foil package and for the Western blot workflow in the AWB, the hard plastic part can easily be removed.

Description

Here it is described how to remove the hard plastic part and substitute it with a similar plastic card with only one large sample well, place it onto the gel, and into this large well place an IPG DryStrip with a length of for example 7 cm. The gel is then run just as a standard gel in the Amersham WB system, automatically scanned and can be evaluated with dedicated software.

IPG DryStrips or gels have an immobilized pH gradient cast on a supporting plastic backing and are sold dry and frozen, and should be rehydrated in urea-containing solution prior to use. IPG DryStrips are provided in 24, 18, 13, 11, and 7 cm lengths; the 7 cm strips come in 6 different pH-ranges. The overall size of a 7 cm IPG Dry strip is approximately 79 x 3.5 mm with a 70 x 3.5 x 0.5 mm gel.

The first prototype 2D card was made in PMMA (Plexiglass) and had features for keying to the AWB system, and an opening for 7 cm IPG DryStrips (Figure 1). The size of the opening is 82 x 5 mm.

Figure 1. Basic design of the 2D card.

After initial trials, the 2D card was improved to more exactly mimic the hard plastic part of the AWB gel card (Figure 2). In particular, the lower area of the card was made thinner not to exert too much pressure on the...