Description

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Transfection Reagent for HT-29 Cells (Colorectal Adenocarcinoma Cells)

• A nanoparticle-based liposome formulation
• Transfection protocols provided for transfection of proteins, DNA, mRNA, siRNA, shRNA and microRNA
• Produce higher level of recombinant protein expression with minimal disruption of normal cell function
• Generate physiologically relevant data you can trust
• Effective for plasmid DNA/siRNA co-transfection
• Easy-to-use transfection protocol with reproducible results
• Low cytotoxicity
• Download in vitro HT29 transfection protocol: [PDF]
• Download HT29 CRISPR/Cas9 transfection protocol: [PDF]
• Download PowerPoint presentation for HT-29 cells transfection kit: [PPT]
• Developed and manufactured by Altogen Biosystems

Transfection Efficiency:

Reagent exhibits at least 70% transfection efficiency of siRNA delivery. Transfection efficiency was determined by qRT-PCR.

Transfection Protocol and MSDS:

Download Altogen Biosystems Transfection Protocol: [PDF]

Download MSDS: [PDF]

HT-29 Cell Line:

According to the Colon Cancer Alliance (CCA), the incidence rates for colon cancer are on the rise in patients younger than age 50, and new treatment regimens are desperately needed. The tumorigenic HT-29 colon cancer cell line is an important tool for the synthesis and investigation of novel cancer drugs. HT-29 was derived in 1964 from the tumor of a 44-year-old woman with colon adenocarcinoma. HT29 cells are human intestinal epithelial cells, which produce the secretory component of Immunoglobulin A (IgA) and carcinoembryonic antigen (CEA). The HT-29 line is designated heterotransplantable, forming well-differentiated grade I tumors. The structure of HT-29 cells include microvilli, microfilaments, mitochondria, smooth and rough endoplasmic reticulum with free ribosomes, lipid droplets, limited primary lysosomes and many secondary lysosomes. The cells express urokinase receptors but do not have detectable plasminogen activator activity. HT-29 cells have been shown to be negative for CD4, but there is expression of galactose ceramide. HT-29 cells are commonly employed for tumorigenicity studies. Also, the HT-29 cell line is extensively used as model system to study epithelial differentiation in vitro. The HT-29 cell line a suitable host for gastric cancer research as well as for studying human infections related to the stomach. Altogen Biosystems offers nanoparticle-based transfection reagent kits that have shown low cytotoxicity for the HT-29 cell line.

Data:

Figure 1. siRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into HT-29 cells following the recommended protocol. At 48 hours post-transfection the cells were analyzed by qRT-PCR for Lamin A/C 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=4).

Figure 2. Protein expression of Lamin A/C in HT-29 cells. DNA plasmid expressing Lamin A/C or siRNA targeting Lamin A/C were transfected into HT-29 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 HT-29 transfection reagent citation references:

• Clin Cancer Res. 2010 16(16). Inhibition of Hypoxia Inducible Factor-1α by Dihydroxyphenylethanol … Terzuoli et al [PDF]
• Oncogene. 2012 (31). EGFR signaling upregulates expression of microsomal prostaglandin E … Donnini et al [PDF]
• Clinical Cancer Research 2010 Aug 15;16(16):4207-16. Inhibition of hypoxia inducible factor-1alpha by dihydroxyphenylethanol, a product from olive oil, blocks microsomal prostaglandin-E synthase-1/vascular endothelial growth factor expression and reduces tumor angiogenesis. Terzuoli et al [PDF]

Altogen Biosystems:
Altogen Biosystems provides preoptimized transfection products for life science research applications. Transfection protocols are optimized for individual cancer cell lines. Altogen Biosystems developed two types of in vivo delivery kits for animal research: Tissue-targeted reagents (delivery to liver, pancreas, and kidney tissues), and broad range in vivo delivery reagents (PEG-Liposome, Nanoparticle-based, Lipid-based, and Polymer-based kits). Advanced formulation of reagents and optimized transfection protocols provide efficient intracellular delivery of proteins, DNA, RNA, and any other negatively charged molecules in vitro and in vivo. Read more about transfection technology at Altogen’s Transfection Resource.

Altogen Labs Research Services:

Altogen Labs provides GLP-compliant contract research studies for pre-clinical research, IND applications, and drug development. Our biology CRO services include: Xenograft services (over 60 inhouse 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 #1771)
• 1.5 ml (Catalog #1772)
• 1.5 ml CRISPR (Catalog #2157)
• 8.0 ml (Catalog #1773)

Altogen生命科学公司致力于研发，生产和销售特定细胞系的转染试剂，用于细胞间生物分子的传递，并通过对转染试剂类型的设计将siRNA和质粒DNA有效地转入不同的细胞系和原代细胞内。 Altogen公司开发的聚合物，脂质体，纳米粒子为基础的转染技术分别针对分子生物学，组合化学，和细胞生物学而分别应用。 Altogen定制服务提供符合GLP要求定制研究服务，包括代稳定的细胞系，细胞银行和冷冻保存，焦磷酸测序，克隆，RNA干扰（RNAi）和基因沉默服务，发展分析，siRNA文库筛选，并转染服务。稳定的肿瘤细胞株和原代细胞的产生，可以是非常昂贵和费时。该公司的细胞培养科学家的细胞株的选择，无论是利息或shRNA表达载体的稳定表达的基因改造。标准的RNAi技术服务，包括设计与合成的siRNA的利益，验证siRNA的沉默效率，siRNA转染条件的优化，使高效的基因沉默细胞系或原代培养细胞的靶基因。 转染培养细胞的瞬时或稳定的引入外源性分子和遗传物质（即RNA或DNA），通常是在生物实验室用来研究基因功能，基因表达的调节，生化映射，突变分析，和蛋白质的生产。科学家利用各种载体分子，这种分子，使质粒DNA（PDNA），信使RNA（mRNA），短干扰RNA（siRNA），小分子RNA（miRNA）的，并进入肿瘤细胞株和原代细胞的蛋白质的基因交付。不幸的是，无单提货的方法或转染试剂，可以适用于所有类型的细胞，细胞的细胞毒性和转染效率显着不同，取决于试剂，协议，并正在利用细胞类型。 Altogen生物系统公司提供超过60种类型的细胞的预优化转染试剂盒。 纳米粒子，脂质和聚合物基ALTOGEN ®在体内转染试剂，使交付功能的RNA和DNA分子在体内。 PEG脂质体在体内输送系统减少由于PEG修饰的先天免疫反应，并提供高效的siRNA转染的DNA，并在体内的蛋白质。 由科学“杂志（2010年12月17日）：PEG脂质体在体内转染试剂盒siRNA的特色 Altogen生物系统功能的特定细胞系转染试剂盒：   公司：www.altogen.com