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. 2008 Feb 24;99(5):1000–1007. doi: 10.1111/j.1349-7006.2008.00770.x

Figure 5.

Figure 5

Association of cluster of differentiation 98 (CD98) heavy chain (hc) with the antigen recognized by SOL22 and SOL69. (a) HeLa cells were stained with SOL22 (upper) or SOL69 (lower) with species‐specific fluorescein isothiocyanate (FITC)‐conjugated anti‐rat IgG, and simultaneously stained with HBJ127 anti‐CD98 hc with species‐specific TexasRed‐conjugated anti‐mouse IgG. (b) Sandwich‐type enzyme‐linked immunosorbent assay was carried out with a combination of anti‐human CD98 hc human monoclonal antibody (mAb) (HH25) as the capture mAb, and anti‐human CD98 hc mouse mAb (HBJ127), SOL22, or SOL69 as the detecting mAb using HeLa cell lysates. (c) The lysates of biotin‐labeled HeLa cells were immunoprecipitated with SOL22 or anti human CD98hc mAb (4F2), subjected to sodium dodecylsulfate–polyacrylamide gel electrophoresis (SDS‐PAGE), and blotted onto polyvinylidene difluoride (PVDF) membranes, and proteins were visualized using Elite ABC and substrates. (d) The lysates of HeLa cells were immunoprecipitated with SOL22 or 4F2, subjected to SDS‐PAGE, and blotted onto PVDF membrane, and proteins were immunostained with anti‐CD98 mAb (HH25). LAT, l‐type amino‐acid transporter; lc, light chain.