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Fig. 2 | BMC Biotechnology

Fig. 2

From: Secretion of Fc-amidated peptide fusion proteins by Chinese hamster ovary cells

Fig. 2

Characterization of endogenous CHO cell PAM. a Five isoforms of Cricetulus griseus PAM are included in the primary RefSeq assembly of the Chinese hamster genome; these isoforms closely resemble those observed in rat, mouse and human: XP_003505817.1, isoform 1; ERE82825.1, isoform 2; XP_003505819.1, isoform 3; XP_003505820.1, isoform 4; XP_003505821.1, isoform 5. b The specific activity of endogenous PHM in the soluble (TM) and solubilized (TMT) crude particulate fractions is indicated. c The solubilized particulate fraction from two separate preparations of wildtype CHO cells (224 μg protein) and from CHO cells stably expressing PAM1 (20 μg protein) were immunoprecipitated using an affinity-purified antibody to the C-terminus of PAM, eluted and fractionated by SDS-PAGE, transferred to PVDF membranes and visualized using affinity-purified antibody to the PHM domain (JH1761); molecular weights of marker proteins analyzed along with PAM1 are indicated. A lighter exposure is shown for the other Wt sample so that the doublet of 92 and 97 kDa bands is visible. Apparent molecular weights calculated from three separate analyses are shown ± Std Dev. d PHM and PAL assays were performed on lysates prepared from wildtype CHO cells and from CHO cells stably expressing rat PAM1 or rat PAM 820 s; lysates were prepared in 1 % TX-100

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