However, differing spot positions will still occur in any nontrivial experiment that includes more than one gel.
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For example, the membrane protein FTT_0583 was observed to be immunoreactive in two discrete protein spots, differing in both MW and pI (Figure 1a & b).
The other two of the 11 proteins were detected as pairs of spots differing in both molecular weight and isoelectric point (SCO5465, SCO2271).
In some cases, the same immunoreactive protein focused to more than one area on a gel, with protein spots differing slightly in isoelectric point.
Quantitative spot analysis of the young and senescent cell proteome revealed 29 spots differing significantly in abundance (p ≤ 0.05)).
Several cyclophilins, lately related to defense reactions in the Arabidopsis- Pseudomonas system, were also identified in the study and from more than one spot differing slightly in its p I value.
In total, 558 protein spots differed in fluorescent signal intensity by at least 1.5 fold (p ≤ 0.05) between the samples, with 246 spots (13% of total detected) enriched in fibres and 312 spots (18% of total) enriched in the non-fibre fraction.
In addition to the gel-to-gel variations, these normalization methods have to cope with differing backgrounds and differing distributions of spot quantities between dyes.
In the first step, all four MDS subgroups were compared together and spots that were found to significantly differ (ANOVA P < 0.05) among the groups were selected; 42 different spots were found.
On average, there were 520 ± 27 distinct protein spots on each gel and 1.6 ± 0.7% of the spots differed quantitatively in their expression (p ≤ 0.05 and fold change ≥2) between infected B19 and B21 chickens.
