Antibodies Western blotting

FIGURE 10.3 Structure of the BHTOH-KLH conjugate utilized for raising polyclonal antibodies. Western blots from 2D gels of cytosolic proteins isolated from mouse lung epithelial cells. The blot on the right is from cells incubated with BHT-QM and the blot on the left is from untreated cells. Immunoreactive proteins were identified by electrospray LC-MS analysis of the tryptic digests. Source From Ref. 39, with permission from the American Chemical Society. 

DNA, RNA, or protein, which can then be probed with a labeled sequence or antibody (Western blot assays), microscopy embedding, electrotechniques, skin protectants, microfilters, and others. Nitrocellulose continues to be used in photography, the manufacture of lacquers, patent and natural leathers, artificial pearls, process engraving, and cements. 

Western blot A method for transferring protein to a nitrocellulose filter, on which the protein can be detected by a suitable probe (eg, an antibody). 

A. Western blot analysis of membrane fractions from P. vulgaris probed with a PGIP-specific antibody. 1) PGIP purified from P. vulgaris] 2) and 3) membranes 4) supernatant of the membrane preparation. 

Figure 5, Western blot analyses. The blot was probed using antibodies raised against PL3. The PL3 protein bands corresponded to a molecular mass of 42 kDa.

Products were analysed by SDS-PAGE followed by Western blotting (Burnette 1981). RGase antibody production is described by van der Veen et al. 1991 This method can be used to determine the presence of RGase in different enzyme-preparations and culture-filtrates. 

A. nidulans with selected phages of classes A, B, C, D and E resulted in increased polygalacturonase activity in comparison with the wild type strain. The culture media of pga transformants were further analysed by Western blotting with polyclonal antibodies raised against PGI and in all samples examined cross-reactive bands with molecular masses similar to that found for PGI or PGII were detected. 

Vande Woude That was true for both mouse and Xenopus. Even though the protein can be detected by western blot, it is occluded from recognition by antibodies at metaphase II arrest. We speculated that this was somehow uncoupling the interaction with microtubules, which activates some kind of checkpoint. This is the only place that this is seen. If you look at CENPE in somatic cells during M phase, it can always be detected. 

Various methods have been used to examine the composition of proteins adsorbed to SAMs. Overall adsorption patterns can be examined with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) [50, 76, 77]. Absorbed proteins are eluted from the surface with surfactant (SDS), and then separated by electrophoresis. The proteins of interest are examined by western blotting [50, 76, 77]. Protein-specific antibodies can be used to detect proteins of... 

Fig. 8.1 Western blot analysis of transgenic lines showing the expression of an assembled monoclonal antibody in transgenic chloroplasts. Lane 1 Extract from a chloroplast transgenic line, Lane 2 Extract from an untransformed plant. Lane 3 Positive control (human IgA). The gel was run under non-reducing conditions. The antibody was detected with an AP-conjugated goat anti-human kappa antibody.

If the gel separates DNA and the DNA is detected with a DNA probe, it is called a Southern blot. If RNA is separated on the gel and then detected by a DNA probe, it is a Northern. A Western uses specific antibodies to detect specific protein molecules on a blot of a protein gel. In the Western blot, the role of the DNA probe is filled by an antibody that recognizes a specific protein. 

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