Antibody Microarray: A Technique to Study the Protein Expression of miRNA Treated Lung Cancer Cells

Abstract

Source: Hossian, A. et. al. Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis. J. Vis. Exp. (2019).

This video describes the technique to study the protein expression in miRNA transfected lung cancer cells by using antibody microarray to measure specific gene expression. In the featured protocol, we perform antibody microarray for the identification of the activity of a combinatorial miRNA treatment in halting cell cycle and angiogenesis.

Protocol

1. miR-143 and miR-506 transfection

CAUTION: Use latex gloves, protective eyeglasses, and a laboratory coat while performing the described experiments. When required, use the biosafety cabinet with the cabinet fan on, without blocking the airways or disturbing the laminar airflow. Always set the protecting glass window to the appropriate height, as described by the manufacturer.

1. Seed NSCLC A549 cells in a T25 cm² flask/6/96 well plate in DMEM/F12K media supplemented with 10% FBS and 1% penicillin-streptomycin (culture media) in a tissue culture hood and incubate overnight at 37 °C with 5% CO....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
A549 Non Small Cell Lung Cancer Cells	ATCC	ATCC CCL-185
Penicillin-streptomycin 10/10	Antlanta Biologicals	B21210
Fetal Bovine Serum (FBS)	Fisher Scientific	10438026
Antibody Array Assay Kit, 2 Reactions	Full Moon Bio	KAS02
Cell Cycle Antibody Array, 2 Slides	Full Moon Bio	ACC058
ScanArray Express	PerkinElmer		Step 2.33: Microarray analysis software
HyClone Phosphate Buffered Saline (PBS)	Fisher Scientific	SH30256FS
Thermo Scientific Pierce BCA Protein Assay	Thermo Scientific	PI23226
hsa-miR-506-3p miRNA Mimic	ABM	MCH02824
hsa-miR-143-3p miRNA Mimic	ABM	MCH01315