Mitsubishi P91W Instruction Manual download pdf (Page 117)

Imaging Gels

107

Imaging Gels

Use unstained standards with stain-free gels, as some prestained standards

are not detected by the stain-free system. To monitor electrophoresis, use a

1:1 mixture of unstained and prestained standards.

Setting up a protocol for stain-free gels is in many ways the same as setting

up protocols for other applications. Follow the instructions in Creating a

Single-Channel Protocol, on page 24, but after step 3, set the activation

options for the gel as follows.

Set the activation options: Visualizing proteins requires activation of the gel.

Choose an activation time option based on your

sample and the purpose of your experiment:

• Use 2.5 min activation when samples are

abundant and when a fully optimized signal-

to-noise ratio is not necessary

• Use 5.0 min activation for detection of proteins

that are in low concentration and for the best

quantification of the maximum number of

bands. Because the reaction is near completion after five minutes,

this method offers an optimal signal-to-noise ratio

Note: If the gel has already been activated for 2.5 min, activating

it for another 2.5 min may improve it; but activating an image for

more than 5 min will not.

Continue setting up the protocol at step 7 under Application Options, on

page 24.

Imaging Blots

To blot stain-free gels, use standard blotting procedures as described in the

instruction manual you are using. Use only PDVF membranes with low

background fluorescence, as membranes other than low fluorescence PDVF

can result in high background or low sensitivity with the ChemiDoc MP

imager.

To assess transfer efficiency, be sure to activate and visualize the gel using the

ChemiDoc MP imager before transfer.

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Mitsubishi P91W Instruction Manual Page 117