Immunoblotting analysis of signaling proteins

Immunoblotting was performed as previously described [16 (link), 18 (link)]. Primary antibodies were obtained from Santa Cruz Biotechnology: α tubulin, 1:10000, sc-23948; BiP, 1:1000, sc-13968; XBP1, 1:1000, sc-7160; Akt, 1:5000, sc-8312; Cell Signaling Technology: pAkt, 1:5000, S473 or Abcam: IRE, 1:2000, ab37073; pIRE, 1:1000, ab48187; TNFα 1:5000 ab66579. Chemiluminescence was performed with Luminata Forte Western HRP Substrate (Millipore) and detected using Chemidoc MP Imaging system. Protein expression was quantified using ImageLab 5.1 software.

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Zaborska K.E., Lee S.A., Garribay D., Cha E, & Cummings B.P. (2018). Deoxycholic acid supplementation impairs glucose homeostasis in mice. PLoS ONE, 13(7), e0200908.

Publication 2018

α tubulin pAkt ab37073 ab48187 ab66579 Luminata Forte Western HRP Substrate

Antibodies Chemiluminescence Protein Tnfα Xbp1 α tubulin

Corresponding Organization : Cornell University

Top 5 similar protocols

Variable analysis

independent variables

Primary antibodies

dependent variables

Protein expression

control variables

Immunoblotting protocol described in previous studies [16, 18]
Chemiluminescence detection with Luminata Forte Western HRP Substrate
Protein quantification using ImageLab 5.1 software

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

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