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Spots number 10, 12, 13 and 24 were found in 2 Western blots, spots 1 and 5 in 3, spot 2 in 4, spot 3 in 5 and spots 4 and 9 in 6.
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The use of multiple sera per Western blot is required to expose several antigen spots per blot which is essential for spot pattern recognition and alignment of silver-stained proteins with Western blot spots.
Two of the nitrocellulose filters were incubated successively with 8 different patient sera each, so to grasp as many antigens as possible and to guarantee that every Western blot displayed several antigen spots which is essential for spot pattern recognition and alignment of silver-stained proteins with Western blot spots.
The serum combinations for multiple probing were selected to allow good pattern definition for assigning Western blot spots with protein spots in the silver-stained gel, and are listed in Table 1 where the numbering of the sera corresponds to the numbering in Figure 1.
Figure 1 d) shows there was no cross-reactivity of secondary antibody to further confirm the positive Western blot spots in Figure 1(c).
Figure 1(c) shows the Western blot image with the labeled positive phosphotyrosine-immunoreactivity, 51 phosphotyrosine immunopositive Western blot spots were detected, and the corresponding silver-stained protein spots were labeled in Figure 1(a).
Timing of HIV acquisition was assessed by RNA or DNA PCR testing on Dry Blot Spot or serum samples from that visit and from prior visits until a negative PCR test was obtained.
Based on its isoelectric point and its relation to the other HP1β isoforms detected in the Western blot, spot 1 in Figure 3E represents most likely the unphosphorylated form of HP1β.
The 2D gel spots corresponding to the phosphotyrosine-positive Western blot spot were excised, and the proteins that were contained in 2D gel spots were digested in gel with trypsin [ 48].
There is no denying the blemishes, blots, and spots that pepper her work, but do they invalidate it, and coarsen its effect?
In highly resolving 2-dimensional Western blots 109 antigen spots were detected with sera of 19 patients, i.e. an average of 5 to 6 per patient.
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