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Miscellaneous Protocols

This section contains some miscellaneous methods that didn't have a home elsewhere.

Protein cleavage includes a method for cleaving proteins with hydroxylamine.

Isoelectric focusing is a procedure specifically adapted to membrane proteins.

CNBr is the procedure we use to couple proteins to CNBr Sepharose to make affinity columns.


GENERAL LAB PROCEDURES


PROTEIN CLEAVAGE

Hydroxylamine Cleavage of Asn-Gly


ISOELECTRIC FOCUSING

Isoelectric Focusing (IEF) of Membrane Proteins


TCA PRECIPITATION OF PROTEINS


CNBr

Coupling protein to CNBr activated Sepharose Beads


TRICINE SDS GELS

Preparation of Stock Solutions Required for Making Tricine SDS Gels

Preparation of Tricine SDS Gels

Preparation of the Stock Solutions Required for Running Tricine SDS Gels

Preparing to Run Tricine SDS Gels


QUANTITATIVE IMMUNOBLOT

Western Blotting

Preparing Samples for Western Blotting

Preparing Samples for Quantitative Western Blotting

Tricine SDS Gels

Electrophoretic separation

Electrophoretic Separation for Quantitative Western Blotting

Transfer

Quantitative Western Blotting

Developing Western Blots Using the ECL Method

Developing Western Blots with Scientific Imaging Film

Visualization of Western Blots on the Kodak Image Station



Please feel free to try any of our protocols.  They are available in web and Adobe Acrobat format.  All that we ask that you credit us (or the original author) when using them.  This document is in a permanent state of flux - no protocol is ever really perfect and therefore it is never finished!