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Cignal Finder Reporter Arrays

用于研究信号通路对基因修饰或化学处理的应答

  • 一次实验可分析多条通路
  • 规格灵活
  • 研究基因敲除和过表达
  • 研究化学调控因子

Cignal Finder Reporter Array可对10条信号通路进行全面的分析。通过同时筛选通路活性,可快速找到相关的通路用于进一步研究。Cignal Finder Reporter Arrays可快速定位特定基因或药物影响的通路。

产品 Product no. 货号 目录价:
 
 
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Available Products
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产品 Product no. 货号 目录价:
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-001L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-003L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-005L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-006L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-007L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
show details
336821 CCA-008L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
show details
336821 CCA-009L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
show details
336821 CCA-010L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
show details
336821 CCA-101L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-103L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-105L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-106L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-107L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-108L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-109L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-110L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-901L varies
200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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336821 CCA-999L varies
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产品 Product no. 货号 目录价:
 
 
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Cignal Finder Reporter Arrays适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


0
p53基因敲除影响信号传导通路。
HEK-293H细胞转染p53 siRNA或阴性对照siRNA,同时共转染来自Cancer 10-Pathway Reporter Array的通路特异报告载体和阴性对照。转染16小时后,将培养基换成完全培养基。转染48小时后,进行双荧光素酶分析。结果表明基因表达变化显著。
1
TNFα影响信号传导通路。
HeLa细胞选用Immune Response 10-Pathway Cignal Finder Reporter Array反转染。转染16小时后,将培养基换成分析培养基。转染32小时后,细胞用5 ng/ml TNFα处理,或不未处理。TNFα处理6小时,进行双荧光素酶分析。结果表明基因表达变化显著。
2
Cignal Finder Array的反应管实验流程。
3
Cignal Finder Array的反应孔板实验流程。
4
确定最佳用药量。
HepG2细胞选用Stress and Toxicity 10-Pathway Reporter Array反转染通路特异报告载体以及阴性和阳性对照。转染16小时后,将培养基换成完全培养基。30小时后,细胞用剂量递增的衣霉素(0–4 µg/ml)处理。48小时后,进行双荧光素酶分析。结果表明基因表达变化显著。0.125 µg/ml衣霉素是抑制糖蛋白合成的最低有效量。
Performance

Cignal Finder Reporter Arrays可用于很多应用。

确定最佳药物剂量

在药物研发过程中,毒性是候选药物失败的主要原因。鉴定特定浓度或化合物的潜在毒性和细胞应答反应,对于今后的研究大有益处。有毒化合物或剂量会激活不同的应激反应通路。基于细胞的荧光素酶报告基因检测采用96孔板的形式,可用于检测10条应激反应通路。

衣霉素抑制GlcNAc phosphotransferase(GPT)酶,引起内质网应急反应。研究中,Stress and Toxicity 10-Pathway Reporter Array用于确定衣霉素的最低有效浓度,而不会激活胞内应急通路。结果表明,0.125 µg/ml衣霉素是抑制糖蛋白合成的最低有效浓度(参见"Identification of optimal drug dose")。

癌症信号通路缺失的细胞应答

研究最成熟的肿瘤抑制基因是p53基因。为了对p53的生物功能有更多了解,检测受p53敲除影响的信号通路非常重要。Cancer 10-Pathway Reporter Array可帮助鉴定受p53敲除调控的关键癌症信号通路(参见"Signaling pathways affected by p53 knockdown")。

结果表明,在HEK-293H细胞中,p53基因的敲除会下调p53信号,而上调Notch、缺氧和MAPK/ERK信号。Notch信号通常在人类恶性肿瘤中下调。p53 RNA干扰激活Noctch信号通路,表明Notch可能是一种原癌基因。

检测细胞因子相关的信号通路

肿瘤坏死因子α(TNFα)是多效炎症因子。研究TNFα调控的重要免疫信号通路非常重要。Immune Response 10-Pathway Cignal Finder Reporter Array能为研究TNFα生物应答相关的信号通路提供有用的信息。

Immune Response 10-Pathway Cignal Finder Reporter Array发现TNFα能激活HeLa细胞中的NFkB和MAPK/JNK信号通路(参见"Signaling pathways affected by TNFα")。

Principle

不论是转染即用型管规格,还是孔板规格,每个芯片都包含10个Cignal Reporter Assays和2个对照。所有的报告基因检测都采用双荧光素酶技术。每个报告系统都是两种载体的混合物,一种是通路相关转录因子应答的萤火虫荧光素酶载体,一种是持续表达的海肾荧光素酶载体。

每个转染子的双荧光素酶结果都计算在内。通过比较处理与未处理转染子报告基因的标准化荧光素酶活性,可决定每个信号通路的活性变化。

相同处理的阴性对照作为特异性对照。阳性对照通过检测GFP表达情况,可作为转染效率对照,或检测萤火虫和海肾荧光素酶作为阳性对照。

Procedure

操作流程包括3个简单的步骤:

转染Cignal Reporter Assays,检测核酸进入细胞 
使用siRNA、蛋白、多肽或目标小分子进行处理 
使用荧光素酶活性检测对报告基因定量分析
Cignal Finder Reporter Array:管规格

Cignal Finder Reporter Arrays管规格采用12联管的形式,含有阳性和阴性对照。可即用于将报告基因系统转染或逆转染进入目的细胞株(参见"Cignal Finder Array, tube format procedure")。

Cignal Finder Reporter Array:孔板规格

Cignal Finder Reporter Array孔板规格采用96孔板形式。每个报告基因和对照检测都在孔板的单个孔中进行(每个检测含8个孔,参见"Cignal Finder Array, plate format procedure")。

Applications

Cignal Reporter Arrays非常适合研究表型相关的RNAi或过表达实验,小分子或化合物的生物应答,以及蛋白、多肽和配体的作用机理。

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