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This finding is supported by immunostaining of single muscle fibers for LAMP-1 which shows remnants of lysosomal membranes (Figure 8).
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Three patients - D4, D12, and D13 (currently aged 66, 65, and 48, respectively) - showed minimal or no pathology by both immunostaining of single muscle fibers and routine histological examination of muscle sections (Table 1).
We conclude that antibody-conjugated COINs bind specifically to antigens when used for immunostaining of single cells.
Such manipulations are followed by immunostaining of membrane proteins, followed by automated imaging to quantify single-cell fluorescence signals.
This was further confirmed by immunostaining of cleaved caspase 3, a marker of programmed cell death (Fig. 3b, c).
We confirmed this suppressor effect by immunostaining of iNOS protein using automated cytometry (Fig. 4g) and microscopy (Supplementary Fig. S4).
Typically, a serum demonstrated immunostaining of a single protein band on the blot.
The specificity of the immunostaining was checked by omission of single steps in the protocol (negative control), by replacement of the primary antibody with preimmune serum and peptide competition tests.
We further analyzed the ability of DSB repair when MS2-CtIP was overexpressed, by performing immunostaining of γ-H2AX staining.
For p53, this tends to reflect adequately the expression of mutant p53 protein, as we have shown in the past by comparing p53 immunostaining and the detection of point mutations by means of single-strand conformation polymorphism [ 10].
Some cells only had single immunostaining of either protein (results not shown for non-specific control sections).
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