User Manual

Overlay Agarose. One bottle containing 50 ml of 0.5% low melting point agarose in 25 mM

Tris, 192 mM glycine, 0.1% SDS, and a trace of Bromophenol Blue.

Instruction Manual. One.

Note:

CHAPS is 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, a

zwitterionic detergent.

Bio-Lyte

3/10 ampholytes is a mixture of carrier ampholytes, pH 3–pH 10.

Other Materials Needed

• PROTEAN IEF cell

• ReadyStrip IPG strips, pH 4-7 (7 cm, 11 cm, or 17 cm)

• IEF focusing tray with lid (same size as IPG strips)

• Electrode wicks, precut

• Blotting filter papers

• Mineral oil

• Forceps

• Pipets for volumes ranging from 4-1000 µl

• Stirplate

• Plastic (Saran) wrap

• 8-16% SDS-PAGE gels (Ready Gel

, Criterion, or PROTEAN II Ready Gel precast gels

• SDS-PAGE electrophoresis cell (Mini-PROTEAN

3, Ready Gel, Criterion, or PROTEAN II XL cell)

• Power supply appropriate for SDS-PAGE system

• Tris/glycine/SDS running buffer

• SDS-PAGE protein stain (Bio-Safe Coomassie or Coomassie Brilliant Blue R-250)

• IEF protein stain (IEF stain or Bio-Safe Coomassie)

• Destain solution (40% methanol, 10% acetic acid)

• Disposable rehydration/equilibration trays with lid (same size as IPG strips)

• SDS-PAGE gel staining trays

• 100 ml graduated cylinders

• High-purity water

Section 3

Storage

Store the entire kit at 4ºC.