Criterion Stain Free™ Gel Imaging System

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Criterion Stain Free™ Gel Imaging System


Question:

Can Criterion Stain Free™ gels be used with blotting applications?


Answer:

Criterion Stain Free gels are compatible with western blotting and provide a quick assessment of protein-transfer efficiency. After blotting, both the gel and the blot can be imaged to assess how much protein remains in the gel and how much was transferred to the membrane. There is no need for post-transfer staining. The images give quantitative data detailing any uneven transfer across the blot, which can then be used to achieve more precise quantitative immunodetection.

The fluorescent staining of proteins in Criterion Stain Free gels is the product of a reaction (activation) with tryptophan (Trp) residues in the proteins. Since Trp is only about 1–2% of the residues in most proteins, most antibodies will not be affected by the Trp modification. A chemiluminescent signal may have a slightly reduced band intensity but the proteins are still detected.

Polyclonal antibody detection is not affected by activation. Detection using monoclonal antibodies may be unaffected or may show decreased signal if the epitopes of the sample protein contain Trp residues. If the epitopes do not contain Trp, there should be no effect on the binding of monoclonal antibodies.

For more information on blotting applications, see Stain-Free Approach to Western Blotting Applications, bulletin #RP0051.


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