Protocol detail

Western Blot Antibody Optimization

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Western blot analysis was performed under conventional conditions. The antibodies used were as follows: a rabbit anti-human S100A11 antibody that we made17–19 (link), mouse anti-human S100A11 antibody (MBL, Nagoya, Japan), rabbit anti-S100A6 antibody (Cell Signaling Technology, Beverly, MA, USA), rabbit anti-calgranulin A (S100A8) antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-calgranulin B (S100A9) antibody (Santa Cruz Biotechnology), mouse anti-RAGE antibody (R&D Systems), mouse anti-tubulin antibody (Sigma-Aldrich, St. Louis, MO, USA), mouse anti-fibroblast antibody (EMD Millipore, Temecula, CA, USA), mouse anti-α-smooth muscle actin antibody (DAKO Agilent Technologies, Santa Clara, CA, USA), rabbit anti-phospho-p70 S6 kinase antibody (Cell Signaling Technology), rabbit anti-p70 S6 kinase antibody (Cell Signaling Technology), rabbit anti-phospho-S6 antibody (Cell Signaling Technology), mouse anti-S6 antibody (Cell Signaling Technology), mouse anti-HA antibody (Cell Signaling Technology), and goat anti-GST antibody (GE Healthcare Bio-Sciences, Piscataway, NJ, USA).

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Takamatsu H., Yamamoto K.I., Tomonobu N., Murata H., Inoue Y., Yamauchi A., Sumardika I.W., Chen Y., Kinoshita R., Yamamura M., Fujiwara H., Mitsui Y., Araki K., Futami J., Saito K., Iioka H., Ruma I.M., Putranto E.W., Nishibori M., Kondo E., Yamamoto Y., Toyooka S, & Sakaguchi M. (2019). Extracellular S100A11 Plays a Critical Role in Spread of the Fibroblast Population in Pancreatic Cancers. Oncology Research, 27(6), 713-727.

Publication 2019

mouse anti-tubulin antibody rabbit anti-p70 S6 kinase antibody rabbit anti-phospho-S6 antibody mouse anti-S6 antibody mouse anti-HA antibody goat anti-GST antibody

Anti antibody Antibodies Antibody Calgranulin a Calgranulin b Fibroblast Goat Human Mouse Mouse α smooth muscle actin P70 s6 kinase Rabbit Rage Tubulin Western blot analysis

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Variable analysis

independent variables

Antibodies used for Western blot analysis

dependent variables

Protein expression levels of S100A11, S100A6, S100A8, S100A9, RAGE, tubulin, fibroblast, α-smooth muscle actin, phospho-p70 S6 kinase, p70 S6 kinase, phospho-S6, and S6

control variables

Western blot analysis was performed under conventional conditions

controls

Positive controls: None specified
Negative controls: None specified

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