Western Blotting Transfer Products

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Introduction

In order to visualize your target protein through antibody binding, the separated protein sample produced by gel electrophoresis must be transferred to a membrane. Using an electric field, the proteins in the PAGE gel can be pushed out of the gel matrix and onto a nitrocellulose membrane while maintaining their position in separate bands. This exposes them on the surface of the membrane, allowing effective incubation, washing, and substrate development, unimpeded by the matrix of the PAGE gel.

Procedure

After soaking your membrane in methanol, immerse it in Boster transfer buffer until it become saturated and sinks. Assemble the transfer cassette from the bottom up, starting with the cathode plate, and building up in the following order: buffer-soaked Boster transfer pad, filter paper, gel, membrane, filter paper, Boster transfer pad, anode plate. Take great care to avoid trapping any bubbles between the layers, and avoid moving the membrane after it has contacted the gel.

Run the transfer setup for approximately 30 minutes at 25V.

For a detailed protocol for membrane transfer, see our Western blot handbook.

Tips

  1. Optimize the transfer type to get the best results for your application
  2. To avoid ghost bands, streaks, and splotches, take care to avoid trapped bubbles between the membrane and gel, and do not move the membrane after it has contacted the gel. See our troubleshooting guides for more information.
Pack Size:0.22um, 9cm
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Pack Size:12.5cm

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