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The ECL kit (Amersham) was used to visualize the protein bands transferred from the blot to film.
Figure 1 demonstrates that the kit was able to detect as per the positive criteria to a tissue-ratio of 1.19×102 mg−/mg+, where all glycoform bands were clearly distinguishable from the blot background.
This difference was markedly enhanced after 72 hours incubation, i.e. the ZO-1 expression in cells treated with 2000 ng/mL DON from basolateral disappeared completely from the blot (Fig. 6).
Stained bands were cut from the blot membrane and subjected to Edman degradation in a Procise sequencer (Perkin Elmer Corp).
In supershift experiments using an antibody against phosphorylated CREB this band disappeared from the blot because it was shifted to much higher molecular weight.
The WWOX probe was stripped from the blot and the membrane was re-hybridised with the human β-actin cDNA control probe.
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g Relative expression of Mecp2 variants on colon epithelium measured from the blots showed in f (n = 3).
To address these issues, the scientific community is now adopting the use of total lane density from the blot-transferred protein as a means of data normalization [ 24– 26].
We observed a disappearance of the radioactive signal from the blots at pH of 14.0.
The Nitrocellulose membranes were removed from the blotting apparatus and dried completely at room temperature.
Representative images chosen for Western blot were non-adjacent bands from the same blot.
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