Western Blot Analysis of Cell Signaling

Western blot was performed as previously described [21 (link)]. Primary antibodies, anti-IRE1α, anti-HSL, anti-p-HSL^Ser660, anti-eIF2α, anti-p-eIF2α^Ser51, anti-BiP and anti-p-PKA substrates (all from Cell Signaling Technology, MA, USA), anti-UCP1 and anti-ATF4 (Santa Cruz Biotechnology) and anti-β-actin (Sigma, MO, USA) were incubated at 4°C overnight, and specific proteins were visualized by ECL Plus (GE Healthcare, Buckinghamshire, UK).

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Xiao Y., Xia T., Yu J., Deng Y., Liu H., Liu B., Chen S., Liu Y, & Guo F. (2016). Knockout of inositol-requiring enzyme 1α in pro-opiomelanocortin neurons decreases fat mass via increasing energy expenditure. Open Biology, 6(8), 160131.

Publication 2016

anti-IRE1α anti-HSL anti-p-HSLSer660 anti-eIF2α anti-p-eIF2αSer51 anti-BiP anti-β-actin ECL Plus

Actin Antibodies Atf4 Ire1α Proteins Ucp1 Western blot

Corresponding Organization :

Other organizations : University of Chinese Academy of Sciences

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Variable analysis

independent variables

Primary antibodies: anti-IRE1α, anti-HSL, anti-p-HSL^Ser660, anti-eIF2α, anti-p-eIF2α^Ser51, anti-BiP and anti-p-PKA substrates (all from Cell Signaling Technology, MA, USA), anti-UCP1 and anti-ATF4 (Santa Cruz Biotechnology) and anti-β-actin (Sigma, MO, USA)

dependent variables

Specific proteins visualized by ECL Plus (GE Healthcare, Buckinghamshire, UK)

control variables

Incubation of primary antibodies at 4°C overnight

controls

No positive or negative controls explicitly mentioned

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