Western Blot Quantification of Signaling Proteins

Western blot analysis was performed as previously described [26 (link)]. The primary antibodies used in this study were anti-CPNE1 (ab155675) (Abcam, UK), anti-RACK1 (sc-17754) (Santa Cruz, USA), anti-pEGFR (Tyr1068) (1H12) (#3777), anti-EGFR (#4267), anti-pAKT (Ser473) (D9E) (#4060), anti-AKT (#4685), anti-pERK (Thr202/Tyr202) (D13.14.4E) (#4370), anti-ERK (#4695), and anti-β-actin (#3700) (Cell Signaling Technology, USA). The bands were developed by an electrochemiluminescence reagent, imaged with a ChemiDoc XRS + (Bio-Rad, USA), and finally quantified with ImageJ software (National Institutes of Health, Bethesda, MD, USA).

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Wang A., Yang W., Li Y., Zhang Y., Zhou J., Zhang R., Zhang W., Zhu J., Zeng Y., Liu Z, & Huang J.A. (2022). CPNE1 promotes non-small cell lung cancer progression by interacting with RACK1 via the MET signaling pathway. Cell Communication and Signaling : CCS, 20, 16.

Publication 2022

anti-RACK1 (sc-17754) anti-AKT anti-ERK anti-β-actin ChemiDoc XRS +

Actin Antibodies Egfr Western blot analysis

Corresponding Organization :

Other organizations : Soochow University, First Affiliated Hospital of Soochow University

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Variable analysis

independent variables

Primary antibodies used in this study

dependent variables

Protein expression levels of CPNE1, RACK1, pEGFR, EGFR, pAKT, AKT, pERK, ERK, and β-actin

control variables

Western blot analysis protocol as previously described [26]
Electrochemiluminescence reagent for band development
ChemiDoc XRS+ for imaging
ImageJ software for quantification

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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