サイトカイン & 成長因子 – 品質
サイトカイン製品の品質
最高性能のサイトカインを提供するために、厳密な品質管理基準を設けています:
- ほとんどの製品にタグや余計なアミノ酸がなく、多くを哺乳類発現系で産生することで、天然に近い立体構造と翻訳後修飾を実現しています。
- ロット毎に、関連する細胞を用いたセルベースアッセイでED50 を決定しています。
- SDS-PAGE により、ほとんどのサイトカインが98% 以上の純度であることを確認しています。
- 生物活性と純度にばらつきがないように、複数ロットの相対比較試験を実施しています。
- エンドトキシン濃度をLAL アッセイで測定し、0.01 ng/μg 以下であることを確認しています。
- SDS-PAGE の純度検定には還元タンパク質を、ジスルフィド結合による二量体の確認には非還元タンパク質を使用しています。
- 生物活性と純度のデータは、全ての製品の製品ページとデータシートに掲載しています。
Purity
The purity of recombinant Human Vascular Endothelial Growth Factor-121 (hVEGF121) #8908 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hVEGF121 and staining overnight with Coomassie Blue. Heterogeneity in SDS-PAGE is due to glycosylation. The non-reduced cystine-linked homodimer migrates as a 30–36 kDa protein.
Bioactivity
The proliferation of HUVEC treated with increasing concentrations of Human Vascular Endothelial Growth Factor-121 (hVEGF121) #8908 was assessed. After 72-hour treatment with hVEGF121 cells were incubated with a tetrazolium salt and the OD450–OD650 was determined.
Western Blot
Western blot analysis of extracts from HUVEC untreated or treated with Human Vascular Endothelial Growth Factor-121 (hVEGF121) #8908 for 15 minutes, using Phospho-p38 MAPK (Thr180/Tyr182) (3D7) Rabbit mAb #9215 (upper) and p38 MAPK Antibody #9212 (lower).
Human Tumor Necrosis Factor-α (hTNF-α)
HeLa cells were serum-starved and then treated with increasing doses of Human Tumor Necrosis Factor-α (hTNF-α) #8902 for 20 minutes. IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814 and Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb #3033 were used to assess NF-κB activation. With increasing concentrations of hTNF-α, a decrease in total IκBα (3-fold) and increase in phospho-NF-κB (Ser536) protein (5-fold) was observed, consistent with TNF-α induced degradation of IκBα resulting in activation of NF-κB. The signal for each antibody was analyzed using an Acumen® Explorer and images were acquired with Cellomics ArrayScan® VTI.
Inset: Signal obtained for IκBα, untreated or treated for 20 minutes with 100 ng/ml TNF-α.
The purity of recombinant Human Tumor Necrosis Factor-α (hTNF-α) #8902 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hTNF-α and staining overnight with Coomassie Blue.
The viability of L-929 cells treated with increasing amounts of Human Tumor Necrosis Factor-α (hTNF-α) #8902 in the presence of 2 ng/ml actinomycin D was determined. Cells were stained with crystal violet at the end of treatment and the OD595 was determined.