Western Blot Analysis of TIAM2 and EGFR Signaling

The WB was performed as described before [86 (link)]. Proteins of target cells were extracted using a lysate buffer (#87787, Thermo Fisher) containing a protease inhibitor (#P1010, Beyotime). After the protein concentration was determined by BCA assay (#E112-01, Vazyme), 40 μg was used for WB analysis. We purchased the rabbit antibody anti-human TIAM2 (#ab199426, Abcam), mouse monoclonal antibody anti-human β-actin (#AF7018, Affinity), rabbit antibody anti-human p-EGFR (Tyr 1068) (#AF3045, Affinity), rabbit antibody anti-human EGFR (#AF6043, Affinity), rabbit antibody anti-human AKT1/2/3 (Ser473) (#AF0016, Affinity), rabbit antibody anti-human AKT (#AF6261, Affinity), goat anti-rabbit antibody (#S0001, Affinity), and goat anti-mouse antibody (#A21010, Affinity).

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Liang L., He H., Jiang S., Liu Y., Huang J., Sun X., Li Y., Jiang Y, & Cong L. (2022). TIAM2 Contributes to Osimertinib Resistance, Cell Motility, and Tumor-Associated Macrophage M2-like Polarization in Lung Adenocarcinoma. International Journal of Molecular Sciences, 23(18), 10415.

Publication 2022

P1010 ab199426

Actin Akt1 Anti antibody Antibody Assay Buffer Cells Egfr Goat Human Monoclonal antibody Mouse Protease inhibitor Protein Proteins target Rabbit Tiam2

Corresponding Organization : Hunan Normal University

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Variable analysis

independent variables

Protein extraction method using lysate buffer containing protease inhibitor

dependent variables

Expression levels of TIAM2, β-actin, p-EGFR, EGFR, AKT1/2/3 (Ser473), and AKT

control variables

Protein concentration (40 μg used for Western blot analysis)
Primary antibodies: anti-TIAM2, anti-β-actin, anti-p-EGFR, anti-EGFR, anti-AKT1/2/3 (Ser473), anti-AKT
Secondary antibodies: anti-rabbit, anti-mouse

controls

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

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