Protein Analysis in Colorectal Cancer

The preparation of cell extracts for protein analyses, immunoprecipitation and Western blotting procedures have been described [38 (link), 39 (link)]. The preparation of the cell extracts from the paired samples (normal and tumor tissue) from patients with CRC were performed as described below in the xenograft studies. Depending on the molecular weight of the proteins to be analyzed, calnexin, tubulin, or GAPDH were used as loading controls. CA2 was used as a colon normal tissue control [40 (link), 41 (link)]. Densitometric measurements of the bands were performed using the Image Lab™ Software Version 6.0.1 Bio-Rad Laboratories (Hercules, CA, USA), which was provided with a ChemiDoc apparatus. Stain free blot was performed by adding 50 µl of 2,2,2-Trichloroethanol to 10 ml of the SDS-PAGE gel solution. Detection of total protein was made in the ChemiDoc apparatus (Bio-Rad) following the manufacturer's instructions.

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Montero J.C., del Carmen S., Abad M., Sayagués J.M., Barbáchano A., Fernández-Barral A., Muñoz A, & Pandiella A. (2023). An amino acid transporter subunit as an antibody–drug conjugate target in colorectal cancer. Journal of Experimental & Clinical Cancer Research : CR, 42, 200.

Publication 2023

Image Lab™ Software Version 6.0.1 ChemiDoc apparatus

Calnexin Colon Densitometric Gapdh Immunoprecipitation Normal tissue Patients Protein Sds page Stain Trichloroethanol Tubulin Tumor Xenograft

Corresponding Organization :

Other organizations : Universidad de Salamanca, Instituto de Investigación Biomédica de Salamanca, Centro de Investigación Biomédica en Red de Cáncer, Centro de Investigación del Cáncer, Instituto de Investigaciones Biomédicas Sols-Morreale, Autonomous University of Madrid, Hospital Universitario La Paz

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Variable analysis

independent variables

Preparation of cell extracts from paired samples (normal and tumor tissue) from patients with CRC

dependent variables

Protein analyses
Immunoprecipitation
Western blotting

control variables

Calnexin, tubulin, or GAPDH as loading controls, depending on the molecular weight of the proteins to be analyzed
CA2 as a colon normal tissue control

controls

Positive control: CA2 as a colon normal tissue control
Negative control: Not explicitly mentioned

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