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Transfection Reagent for A549 Cells (Lung Carcinoma Cells, CCL-185)
Two component formulation enhances lipid mediated transfection efficiency
Optimized easy-to-use transfection protocol provided for transfection of siRNA, DNA, mRNA, and microRNA
Kit includes Transfection Enhancer reagent and recommended transfection protocol
High transfection efficacy in the presence of serum
Expand your RNAi application with a reagent optimized for delivery of both siRNA and plasmid
Reproducible transfection results
Works well for standard reverse transfection and high-throughput applications
Download PowerPoint presentation for A549 cells transfection kit: [PPT]
Developed and manufactured by Altogen Biosystems
Reagent exhibits at least 80% transfection efficiency of siRNA delivery. Transfection efficiency was determined by real-time RT-PCR.
Transfection Protocol and MSDS:
Download Altogen Biosystems Transfection Protocol: [PDF]
Download MSDS: [PDF]
A549 Cell Line:
Lung cancer is considered the leading cause of cancer death worldwide, despite a steady decline in the mortality rate resulting from widespread public health awareness campaigns. New advances in lung cancer treatment approaches, as well as immunotherapy and targeted therapies, are essential in treating patients with advanced stage disease. According to Dr. John Heymach, chairman of Thoracic Oncology at MD Anderson, approximately 160,000 people die from lung cancer every year, a number more significant than the next three prominent cancers (colorectal, breast, prostate) combined. The A549 lung cancer cell line has been used to investigate treatments for various forms of lung cancer, in addition to other respiratory illnesses such as asthma. The A549 cell line was derived in 1972 from the carcinomatous lung tissue of a 58-year-old Caucasian male. A549 cells are adenocarcinomic human alveolar basal epithelial cells that are squamous and responsible for the diffusion of substances such as water and electrolytes, across the alveoli of lungs. This cell line a hypotriploid human cell line with the modal chromosome number of 66, occurring in 24 percent of cells. There were also frequent instances of cells with 64, 65, and 67 chromosome counts. A549 cells have been shown to synthesize lecithin with a high percentage of desaturated fatty acids via the cytidine diphosphocholine pathway. These cells are used in vitro model for a type II pulmonary epithelial cell model for drug metabolism. Immunoperoxidase staining shows that A549 cells are positive for keratin. This cell line is typically used to investigate numerous respiratory ailments, such as viral infections that can cause asthma, tissue damage linked to asbestos exposure, and smoking-related emphysema. A549 cells grow as a monolayer and are capable of synthesizing lecithin.
Figure 1. SiRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected following the recommended protocol. At 48 hours post-transfection the A549 cells were analyzed by qRT-PCR for gene expression levels. 18S rRNA levels were used to normalize the Lamin A/C data. Values are normalized to untreated sample. Data are means ± SD (n=3).
Figure 2. Protein expression of Lamin A/C in A549 cells. DNA plasmid expressing Lamin A/C or siRNA targeting Lamin A/C were transfected into A549 cells following Altogen Biosystems transfection protocol. At 72 hours post-transfection the cells were analyzed by Western Blot for protein expression levels (normalized by total protein, 10 µg of total protein loaded per each well). Untreated cells used as a negative control.
Selected citations for A549 transfection reagent used in the following publications:
- Journal of General Virology. 2008 89:1978. Role of retinoic acid inducible gene-I in human … Liao et al [PDF]
- PLoS One. 2012 (7). Pharmacological Inhibition of Microsomal Prostaglandin E Synthase-1 … Finetti et al [PDF]
- Mol Cell Biol. 2013 33(7). SCO2 induces p53-mediated apoptosis by Thr845 phosphorylation … Madan et al [PDF]
Altogen Biosystems provides cell type specific and preoptimized transfection products, elecroporation kits, and targeted in vivo delivery reagents. Advanced formulation of reagents and optimized transfection protocols provide efficient intracellular delivery of protein, DNA, mRNA, shRNA, siRNA and other biomolecules. Read more about transfection technology at Altogen’s Transfection Resource.
Altogen Research Services:
Altogen Labs provides GLP-compliant contract research studies for pre-clinical research, IND applications, and drug development. Biology CRO services include: Xenograft models (30+), development of stable cell lines, ELISA assay development, cell-based and tissue targeted RNAi studies, safety pharm/tox assays, and other studies (visit AltogenLabs.com).
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