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Fig. 5. TNGW1 resided in a subset of GWBs. (A) Mouse monoclonal anti-rTNR antibody 2F11 stained a subset of GWBs that were recognized by the prototype serum 18033. HEp-2 cells were stained using mouse mAb 2F11, which stained a subset of GWBs that were detected by serum 18033 (arrows) but did not recognize other GWBs (arrowheads). Culture medium was used as a negative control. Bar, 10 µm. (B) Mouse anti-rTNR mAb 2F11 stained a subset of GWBs that were recognized by anti-GW182 mAb 4B6. HEp-2 cell staining was performed similarly using anti-rTNR 2F11 (IgG2a) and anti-GW182 4B6 (IgG1) and different fluorochrome-conjugated mouse IgG subclass-specific secondary antibodies to evaluate the distribution of TNGW1 in GW182-positive GWBs. Anti-rTNR 2F11 stained some of the foci detected by 4B6 (arrows) but not others (arrowhead). The second and third row of images show controls that had been incubated with either 4B6 or 2F11 and were stained with both secondary antibodies to demonstrate secondary antibody specificities. The asterisks in the first panel shows nonspecific nucleolar staining detected by the anti-IgG2a secondary antibodies. Bar 10 µm.
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