Description

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Transfection Reagent for Fibroblast Cells (Fibroblast Cell Lines and Primary Fibroblasts)

• Proprietary cationic lipid formulation
• High transfection efficiency of small RNA (siRNA, shRNA, miRNA), mRNA, pDNA
• Effective and robust intracellular delivery
• Kit includes Transfection Enhancer reagent
• Work in the presence of serum
• A proven reagent for establishing stable fibroblast cell lines
• Download Fibroblast transfection protocol: [PDF]
• Download Fibroblast CRISPR/Cas9 transfection protocol: [PDF]
• Download PowerPoint presentation for Fibroblast cells transfection kit: [PPT]
• Developed and manufactured by Altogen Biosystems

Transfection Efficiency:

Reagent exhibits at least 85% transfection efficiency of siRNA delivery. Transfection efficiency was determined by qRT-PCR.

Transfection Protocol and MSDS:

Download Altogen Biosystems Fibroblasts Transfection Protocol: [PDF]

Download MSDS: [PDF]

Fibroblast Cell Lines and Primary Fibroblasts:

Fibroblasts maintain the structure of connective tissues by secreting precursors of the extracellular matrix, primarily a variety of fibers and the ground substance. Fibroblasts and related connective tissues sculpt the significant portion of an organism. Fibroblasts are derived from primitive mesenchyme. Fibroblasts have a branched cytoplasm that surrounds a nucleus with one or two nucleoli. Fibroblasts can migrate slowly over substratum as individual cells (in contrast to epithelial cells). Fibroblasts can show different phenotypes in separate anatomical locations. Active fibroblasts have rough abundant endoplasmic reticulum. Fibroblasts often align in parallel clusters when crowded. Unlike the epithelial cells lining the body structures, fibroblasts do not form flat mono-layers and are not restricted to a basal lamina on one side. Of the total tissue mass in the human body, approximately 45% is made up of connective tissue. Within the cells that make up connective tissue, fibroblasts are the most prolific. Fibroblasts secrete extracellular proteins that help maintain the structure of connective tissue. Fibroblasts are well known for the role they play in wound healing, by synthesizing collagen at the site of an injury. Many studies have successfully demonstrated the ability of healthy fibroblasts to convert into different functional cells. Altogen Biosystems manufactures pre-optimized cationic lipid-based transfection reagent kits suitable for carrying out transfection on various fibroblast cell lines.

Data:

Figure 1. siRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into fibroblast cells following the recommended protocol. At 48 hours post-transfection the 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 Fibroblast cells. DNA plasmid expressing Lamin A/C or siRNA targeting Lamin A/C were transfected into Fibroblast 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 Fibroblast transfection kit citation references:

• Journal of Experimental Medicine. 2015. 212 (7) 1061-1080. The Dishevelled-binding protein CXXC5 negatively regulates cutaneous wound healing. Kim et al [PDF]
• Am J Pathology. 2011. 179(4):1792-806. Defective myofibroblast formation from mesenchymal stem cells in the aging murine heart rescue by activation of the AMPK pathway. Cieslik et al [PDF]
• Rep Biochem Mol Biol. 2015. Apr;3(2):76-81.Expression of TGF-β3 in isolated fibroblasts from foreskin. Mahmoudi et al [PDF]
• The American Journal of Pathology. 2011 179(4). Defective Myofibroblast Formation from Mesenchymal Stem Cells … Cieslik 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 offers GLP-compliant contract research studies for pre-clinical research, IND applications, and drug development. Our biology CRO services include: Xenograft services (over 60 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 #1755)
• 1.5 ml (Catalog #1756)
• 1.5 ml CRISPR (Catalog #2142)
• 8.0 ml (Catalog #1757)

AltogenBiosystems是一家开发和制造用于生命科学研究，药物发现和开发的转染试剂盒的生物技术公司。转染试剂盒针对特定癌细胞系和原代细胞培养进行了优化，可将生物分子有效递送到靶组织中。通过先进的试剂配方和优化的转染方案实现体外（癌细胞系）和体内（动物组织靶向试剂、癌细胞系）递送货物分子，包括质粒DNA，各种类型的RNA（mRNA，siRNA，shRNA，microRNA），蛋白质和小分子研究。 

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

120+细胞转染试剂和活体组织靶向试剂盒制造商AltogenBiosystems是一家生物技术公司，开发和制造用于生命科学研究、药物发现和开发的转染试剂盒。Altogen®体内转染试剂可有效地将生物分子导入靶组织。细胞转染试剂盒针对特定的癌细胞系和原代细胞进行了优化。通过先进的试剂配方和优化的转染方案实现货物分子（DNA、RNA、蛋白质）的高效传递。AltogenBiosystems利用高分子化学、分子和细胞生物学的专业知识，开发了新的体内外给药技术。转染是将外源分子导入培养细胞中，常用于研究基因功能、基因表达调控、生化定位和蛋白质生产。不幸的是，由于细胞毒性和转染效率的差异很大，并且取决于所使用的试剂、方案和细胞类型，因此没有一种单一的传递方法或转染试剂可应用于所有类型的细胞。AltogenBiosystems为120多个癌细胞系和原代细胞类型提供优化的转染试剂盒和电穿孔产品。体内转染试剂可实现组织靶向给药。Altogen的转染试剂盒包括用于体外（癌细胞系）和体内（用于动物研究的组织靶向试剂）转染的转染增强剂试剂和转染复合物冷凝器。Altogen实验室提供符合GLP的实验室合同研究服务。我们的生物CRO服务包括异种移植物的疗效、IND应用的pharm/tox研究和安全性测试、分析开发（ELISA、IC-50、qPCR）、90多个异种移植物动物模型、RNAi和基因沉默服务。Altogen的细胞培养科学家通过在28天内培育出稳定的细胞系，将选择的细胞系转化为稳定表达感兴趣的基因。