Description
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Transfection Reagent for A375 Cells (Melanoma Cells, CRL-1619)
- A nanoparticle-based liposome formulation
- Transfection protocols provided for transfection of proteins, DNA, mRNA, siRNA, shRNA and microRNA
- Transfection Enhancer reagent provided with the kit
- 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 A375 transfection protocol: [PDF]
- Download A375 CRISPR/Cas9 transfection protocol: [PDF]
- Download PowerPoint presentation for A375 cells transfection kit: [PPT]
- Developed and manufactured by Altogen Biosystems
Transfection Efficiency:
Reagent exhibits at least 76% 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]
A375 Cell Line:
Prolonged exposure to ultraviolet (UV) radiation dramatically increases the risk of melanoma, which is among the top 10 most prevalent forms of cancer in the US, and the top 20 in the world. There will be approximately 87,000 new cases of melanoma, in the United States in 2017 alone, according to the Skin Cancer Foundation. The A-375 cell line is a useful melanoma model employed in preclinical research to develop new medicines. A-375 cells were established based on skin cells taken from a 54-year-old female patient who suffered from malignant melanoma (skin cancer). These tumorigenic cells exhibit an epithelial cell morphology and are useful as a stable transfection host. A375 can be helpful for molecular and cell biology research, mainly related to skin cancer.
Data:
Figure 1. siRNAs targeting Lamin A/C mRNA or non-silencing control siRNA were transfected into A375 cell line 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 A375 cells. DNA plasmid expressing Lamin A/C or siRNA targeting Lamin A/C were transfected into A375 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 in vivo transfection product citations (ALTOGEN® IN VIVO Transfection Kits used in the following publications):
- Mol Cell Biol. 2013 33(7). SCO2 induces p53-mediated apoptosis by Thr845 phosphorylation … Madan et al [PDF]
- Hypertension. 2015 65(2):430-9. Neurokinin 3 receptor and phosphocholine transferase… Parchim et al [PDF]
- Gastroenterology. 2011 141(2) Differential type I interferon-mediated autophagic trafficking … Desai et al [PDF]
- PLoS Pathog. 2014 10(10) Exosomes from hepatitis C infected patients transmit HCV … Bukong et al [PDF]
A375 Cells Transfection Reagent
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 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).
Volume Options:
- 0.5 ml (Catalog #1167)
- 1.5 ml (Catalog #1168)
- 1.5 ml CRISPR (Catalog #2102)
- 8.0 ml (Catalog #1169)
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。
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的抗体不容易染色,哪为同仁有这方面的经验?另外基因公司卖的cellsignalingtechnology的抗体要1875元,而中山公司卖的这个抗体只有800多,其中差距在哪里?请赐教
本人对软骨石蜡切片做了MMP-13的免疫组化染色,首次做,无法判断染色部位,请各位帮忙看一下
Bone Mesenchymal Stem Cells 作为一个细胞群体,还没有发现有特定细胞表面marker. 对于那些可以代表自我更新和分化的marker, 也不清楚到底要发现哪一个的表达才能确定该细胞就是BMSC。
目前常用的方法,就是采用培养,colony-forming unit-fibroblasts (CFU-F)这个方法。一般BMSC可以24-48小时贴壁。
流式细胞计数,比如STRO-1,但是一般认为STRO-1阳性的细胞更趋向于造血干细胞,和BMSC简单区别还不是很清楚。
这里有个培养分化的产品
http://www.rndsystems.com/pdf/SC020.pdf
GlucosestarvationcausestranslocationofAMPKβ2tothelysosomeinHEK-293cellsthatisdependentonN-myristoylation.Theexperimentwasperformedinβ2KOcellsasinFig.1c,exceptthatthelysosomalMarkerLAMP1(taggedwithRFP)wasco-expressedwiththewild-typeormutantAMPKβ2.Upperpanelsshowmergedimagesstainedblue(4′,6-diamidino-2-phenylindole(DAPI),nuclei),red(LAMP1,lysosomes)andgreen(AMPKβ2,detectedusingantibodyvalidatedine),incellsincubatedwithorwithoutglucosefor20 min.Lowersmallpanelsaremagnificationsoftheareasindicatedbydashedboxesintheupperpanels,showing(LtoR)redandgreenchannelsandmergedimages.
下面的这段话是图注,图注的意思我明白,但是我想知道merge后的图看什么颜色的荧光,蓝色是细胞核,红色是lysosome(位于胞质),绿色是AMPKβ2,该实验是想观察AMPKβ2是否转位到lysosome上了,如果确实发生了AMPKβ2转位到lysosome上,那么merge后是红色与绿色融合在一起,是吗?融合在一起发什么颜色的光了?
(2)另一方面,使抗原抗体结合更稳定。一般不需要,但对表达较弱的抗原可能有用,4度和37度时分子运动方式不同,前者分子碰撞机率和运动速度小于后者,后者结合更快,但敏感性也提高了并易造成非特异染色。
(3)其实,我更赞同后一种说法,因为我尝试把肝脏或睾丸片子从4度过夜拿出后,直接用PBS洗没发生过脱片现象。事实胜于雄辩!
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