Introduction

MPX™ Multiplexer Blotting System

The LI-COR MPX™ Multiplexer Blotting System was designed for screening multiple samples and multiple targets on a single Western blot. The independent channels and multi-sample features of the MPX eliminate common limitations encountered with antibody cross-reactivity using standard Western blot analysis. The MPX can be used to screen primary antibodies (such as polyclonals, monoclonals, and subclass specific mouse antibodies), secondary antibodies, blocking buffers, and much more. All Western blot workflows that generate a membrane of 7.0 x 8.5 cm can be adapted to the MPX format (Figure 147).

The MPX System conserves precious and costly antibodies via low-volume channel ports (160 µL) that match multi-channel pipette spacing, if desired, for a simplified workflow. The nesting-dovetail design allows linkage of multiple units to process multiple blots.

Four gel combs are designed specifically for use with the MPX System if you choose to pour your own gels. The collection of single-marker combs enables you to select one, two, three, or four samples per gel for electrophoresis and transfer to nitrocellulose or PVDF membranes. Alternatively, pre-cast, single-well gels can be used to generate blots.

Processing is as simple as clamping the blot into the MPX, which creates up to 24 independent channels. The range of usable channels per sample is relative to the comb size—from 24 on a single sample to five per sample using a four-sample comb.

Suggested Materials

All products are available from LICORbio (licor.com/bio) unless otherwise noted.

The following methods offer suggestions for electrophoresis and transfer conditions for use with Odyssey Imaging Systems but are intended only as a supplement to the manufacturer’s instructions. Before proceeding, familiarize yourself with the appropriate user manuals and troubleshooting guidelines. Due to the large number of factors that affect protein transfer efficiency and performance, the scope of this document is limited to the materials listed in this section. Alternate materials may be substituted if desired; however, optimization may be necessary.

Gel Reagents

Poured Gel Combs

Electrophoresis Reagents

Transfer Reagents

Detection Reagents