Description

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Transfection Reagent for SKNAS Cells (Neuroblastoma Cells)

• Proprietary cationic lipids formulation
• A proven reagent for establishing stable cell lines
• Optimized transfection protocols are adapted for use with both standard & reverse transfection methods
• Download in vitro SKNAS transfection protocol: [PDF]
• Download SKNAS CRISPR/Cas9 transfection protocol: [PDF]
• Download PowerPoint presentation for SKNAS cells transfection kit: [PPT]
• Developed and manufactured by Altogen Biosystems

Transfection Efficiency:

Reagent exhibits at least 90% transfection efficiency of siRNA delivery. Transfection efficiency was determined by qRT-PCR.

Transfection Protocol and MSDS:

Download Altogen Biosystems SKNAS Transfection Protocol: [PDF]

Download MSDS: [PDF]

SKNAS Cell Line:

Neuroblastoma originates from embryonic neural crest cells and is the most common extracranial solid tumor, affecting children and leading to poor prognosis due to bone metastasis. According to a 2014 article in the Journal of Bone Oncology, mice inoculated with the SK-N-AS neuroblastoma cell line developed bone metastasis like most human neuroblastoma patients. Neuroblastoma represents nearly 6% of all childhood malignancies, with approximately 700 new cases diagnosed annually in the United States, as per the American Cancer Society. The SK-N-AS cell line is isolated from a bone marrow metastasis of a 6-year old female. SK-N-AS is hyperdiploid human cell line with the modal chromosome number of 47. Normal chromosomes N9 and N22 are single. SK-N-AS are distributed for research purposes only by the Memorial Sloan-Kettering Cancer Center. The SK-N-SH line was derived by J.L. Biedler and is different from SK-N-MC as it displays higher levels of dopamine – beta-hydroxylase as well as a longer doubling time. Also, SK-N-SH is used in cell-mediated cytotoxicity assays as a target cell line. Frataxin-depleted cells display actin network reorganization that can be observed in fibroblasts. The cells were similar to control SK-N-AS cells after Nrf2 staining. It has been shown that treating control SKNAS cells by oligomycin or tBHQ can result in the accumulation of NADH quinone oxidoreductase, glutathione-S-transferase subunit 1 and glutathione reductase.  These cells are known to produce large amounts of IGF-2 and express type 1 IGF (insulin-like growth factor) receptors. SK-N-AS cells can be useful in the study or search for medical treatments for certain types of brain cancer. Altogen Biosystems offers in vitro SK-N-AS transfection reagent for neuroblastoma cells.

Data:

Figure 1. Cyclophilin B silencing efficiency was determined by qRT-PCR in the SKNAS cells transfected by Cyclophilin B siRNA or non-silencing siRNA control following the recommended transfection protocol. Cyclophilin mRNA expression levels were measured 48 hours post-transfection. 18S rRNA levels were used to normalize the Cyclophilin B data. Values are normalized to untreated sample. Data are presented as means ± SD (n=3).

Figure 2. Protein expression of Cyclophilin B in SKNAS cells. DNA plasmid expressing Cyclophilin B or siRNA targeting Cyclophilin B were transfected into SKNAS 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 SK-N-AS transfection reagent citation references:

• Nat Commun. 2016 ;7:12057. Signal transduction controls heterogeneous NF-κB dynamics and target gene expression through cytokine-specific refractory states. Adamson A et al [PDF]
• Endocrine-Related Cancer 2016. 23(8):595-607. Role of protein S in castration-resistant prostate cancer-like cells. Ning et al [PDF]
• RNA. 2010 16(11):2108-19. RNase L releases a small RNA from HCV RNA that refolds … Malathi et al [PDF]

Altogen Biosystems:
Altogen Biosystems transfection and electroporation products for life sciences and cancer research. Transfection kits are developed for individual cancer cell line and transfection protocols are optimized for maximum delivery efficiency. Advanced formulation of reagents and optimized transfection protocols provide efficient intracellular delivery of proteins, DNA, mRNA, shRNA, siRNA, and other negatively charged biomolecules in vitro and in vivo. 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 services (over 60 in-house validated xenograft models), 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).

Volume Options:

• 0.5 ml (Catalog #3259)
• 1.5 ml (Catalog #3260)
• 1.5 ml CRISPR (Catalog #2194)
• 8.0 ml (Catalog #7012)