The expression of the proteins was analyzed by SDS-PAGE. The bacterial pellets were suspended in a loading buffer, heated for 5 minutes at 95°C, and 25 µL of each sample was subjected to 15% SDS-PAGE gel. For the western blot, the samples were separated by the SDS-PAGE and transferred in nitrocellulose membranes by using a liquid transfer system (Bio-Rad). The membranes were blocked with skimmed milk in phosphate buffered saline (PBST) (PBS 1% + Tween 20) and then washed several times with PBST. The membranes were incubated with the conjugated His-tag antibody (Roche) for 1 hour at room temperature and developed by Diaminobenzidine (DAB) solution (Roche, Germany).
Diaminobenzidine dab solution
Manufactured by Roche
Sourced in Germany
Diaminobenzidine (DAB) solution is a laboratory reagent used as a chromogenic substrate in immunohistochemistry and other bioanalytical techniques. It is a versatile tool for the detection and visualization of specific target molecules or proteins in biological samples.
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Lab products found in correlation
2 protocols using diaminobenzidine dab solution
1
SDS-PAGE and Western Blot Analysis
2
Western Blot Analysis of Recombinant Proteins
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The integrity of the purified recombinant proteins was confirmed by Western blot analysis. Western blotting was performed according to the standard protocol [14 (link)] using sera from rabbit immunized against Vibrio cholerae and negative sera (as the negative controls) as the primary antibody at 1 : 100 dilution and HRP-conjugated (horseradish peroxidase) goat anti-rabbit IgG (Abcam, United Kingdom) at 1 : 2500 dilution in 1x TBST buffer (10x: 15 mMNaCl, 10 mMTris-HCl (pH = 7.4), 0.1% Tween 20) as secondary antibody. The reactions were developed by diaminobenzidine (DAB) solution (Roche, Germany).
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