Western Blot for Protein Detection

Western blotting was performed as previously described with minor modifications^{39 (link)}. In brief, whole-cell lysates were prepared in RIPA buffer supplemented with a proteinase-inhibitor cocktail. The lysates were clarified by centrifugation, the supernatant was boiled, and separated by SDS-PAGE. The following antibodies were used for western blots: anti-α-tubulin (Santa Cruz Biotechnology, Santa Cruz, CA, USA, B-5–1–2) and anti-IGFBP2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA, C-18).

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Katayama H., Tamai K., Shibuya R., Nakamura M., Mochizuki M., Yamaguchi K., Kawamura S., Tochigi T., Sato I., Okanishi T., Sakurai K., Fujibuchi W., Arai Y, & Satoh K. (2017). Long non-coding RNA HOTAIR promotes cell migration by upregulating insulin growth factor–binding protein 2 in renal cell carcinoma. Scientific Reports, 7, 12016.

Publication 2017

anti-α-tubulin anti-IGFBP2

Antibodies Buffer Cell Centrifugation Igfbp2 Proteinase a Ripa Sds page Western blots α tubulin

Corresponding Organization :

Other organizations : Miyagi Prefectural Hospital Organization, Kyoto University, Tohoku University

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Not explicitly mentioned

control variables

RIPA buffer supplemented with a proteinase-inhibitor cocktail
Anti-α-tubulin (Santa Cruz Biotechnology, Santa Cruz, CA, USA, B-5–1–2)
Anti-IGFBP2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA, C-18)

controls

Positive control: Not specified
Negative control: Not specified

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