Figure_1.tif (241.09 kB)

Antigenic activity of recombinant autoantigens a. Antigenic activity of PL-7 in various conditions.

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posted on 2014-01-14, 03:18 authored by Ran Nakashima, Yoshitaka Imura, Yuji Hosono, Minae Seto, Akihiro Murakami, Kizuku Watanabe, Tomohiro Handa, Michiaki Mishima, Michito Hirakata, Tsutomu Takeuchi, Keishi Fujio, Kazuhiko Yamamoto, Hitoshi Kohsaka, Yoshinari Takasaki, Noriyuki Enomoto, Takafumi Suda, Kingo Chida, Shu Hisata, Toshihiro Nukiwa, Tsuneyo Mimori

Left, purified recombinant PL-7 was eluted and diluted in PBS and coated on ELISA plates. Middle and Right, purified recombinant PL-7 was eluted in PBS and diluted in 8M urea and 2 × SDS sample buffer, respectively, and then coated onto ELISA plates. b. Five recombinant ARS antigens (His-PL-12, His-EJ, GST-Jo-1, GST-KS, and His-PL-7) were prepared as soluble polypeptides in PBS and their antigenic activity was tested in an ELISA using sera from five patients each containing corresponding autoantibodies (only GST-KS was tested using sera from three patients). Six healthy controls were used in each ELISA. c. Purified recombinant ARS antigens were electrophoresed on SDS-PAGE and transferred to a PVDF membrane followed by immunoblot analysis. CBB; Coomassie Brilliant Blue staining of gels, M; molecular weight marker, HC; healthy control, Lane 1; His-PL-12, Lane 2; His-EJ, Lane 3; GST-Jo-1, Lane 4; GST-KS and Lane 5; His-PL-7.