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Functional Genomics - LightSwitch™

LightSwitch™ Reporter System

everything needed for luciferase reporter assays

アクティブ・モティフ社のLightSwitch™ Luciferase Assay Systemは、哺乳動物生細胞における遺伝子発現制御機構およびクロマチン免疫沈降(ChIP)で検出されたタンパク質結合DNA領域の機能を確実に解析することができるルシフェラーゼアッセイ系です。一般的にルシフェラーゼアッセイは、クローニングおよびDNA調製を行うため時間と労力を多く必要としますが、LightSwtich™ GoClone™コレクション(12,000種類以上のヒト3’UTRおよび18,000種類以上のヒトプロモーター導入済みレポーターベクター)を用いれば、迅速にレポーターアッセイを行うことができます。アッセイ試薬および導入試薬は、プロトコルが最適化されているため、他の手法よりも迅速かつ簡便に実験を行うことができます。また、ポジティブ/ネガティブ・コントロールベクター、GoClone™導入済み細胞株、miRNA類似体/阻害剤、合成応答エレメントコレクションもご用意しています。さらに、ベクター導入、部位特異的変異導入、シグナル伝達経路スクリーニング、配列変異機能解析の受託サービスも行っています。

ご注文方法

1.     下記の「Search the Promoter GoClone Collection」または Search the 3’UTR GoClone Collection」を選択して下さい。移動したページ下記にある「SEARCH BY GENE IDENTIFIER」の中にご希望の遺伝子名を入力し「Promoter」「UTR」「Both」のいずれかのボタンをクリックしてください。ご希望の遺伝子の「Prod ID」「Gene Symbol」が表示されます。

2.     ご注文の際には、「商品コード」「Prod ID」「Gene Symbol」の3つ(空ベクターをご希望の場合は「空ベクターの商品コード」を加えた4つ)が必要になります。

LightSwitch Promoter GoCloneの商品コード:「32001

LightSwitch 3’UTR GoCloneの商品コード:   「32011

3.     1回のご注文につき空ベクターを1個無償で提供しています。以下の4種類の空ベクターの中からお好きなものを1個お選びいただいて、その商品コードをお伝え下さい(必要がなければご注文いただかなくても構いません)。なお下記の空ベクター製品は販売もしています。

品名                                                    包装       商品コード

LightSwitch™ Promoter Reporter Vector                 5 µg        32021    

LightSwitch™ 3´UTR Reporter Vector                     5 µg        32022

LightSwitch™ 5´UTR Reporter Vector                     5 µg        32023

LightSwitch™ LR Enhancer Reporter Vector           5 µg        32024

 

重要事項

すべてのLightSwitchレポーターコンストラクトは最適化されたRenSPルシフェラーゼ遺伝子を含んでおります。したがって、最適な結果を得るためには弊社のLightSwitch Luciferase Assay Reagentsのご使用が必須となります(他社のルシフェラーゼアッセイ試薬はRenSPに対応していません)。また、アッセイを行う際は、適切なポジティブ・ネガティブコントロールベクターとご一緒に行うことを推奨します。

LightSwitch Promoter Control Vectors

LightSwitch 3´UTR Control Vectors

Empty LightSwitch Reporter Vectors

 

Brief summaries for each part of the LightSwitch System are below and include links to more comprehensive information. Additional details on the LightSwitch System can be found at the bottom of this page in the sections titled LightSwitch Luciferase Assay Overview and RenSP Luciferase, which describes the optimized luciferase gene used in all LightSwitch reporter vectors. Or, to search the GoClone™ Collections, click one of these buttons:

Button to jump to the LightSwitch GoClone Search page     Button to jump to the LightSwitch GoClone Search page

LightSwitch™ Promoter Reporter GoClone™ Collection

The Promoter Reporter Collection includes over 18,000 promoters cloned from the human genome and available as transfection-ready GoClone constructs. So, you don't need to spend time and effort cloning the promoters needed for your transcriptional regulation studies. Simply use our Search Tool to find vectors with your promoters of interest, then transfect and assay their activity immediately.

LightSwitch Promoter Reporter vector
LightSwitch™ Promoter
Reporter Vector.

LightSwitch™ 3´UTR Reporter GoClone™ Collection &
miRNA Mimics and Inhibitors

The 3´UTR Reporter Collection includes over 12,000 3´UTRs cloned from the human genome into ready-to-transfect LightSwitch reporter vectors. Combined with our large collection of miRNA Mimics and Inhibitors, you have everything needed to study miRNA-3´UTR interactions, validate miRNA targets, measure RNA stability, translation efficiency and the functional impact of miRNAs on a gene-by-gene basis.

LightSwitch 3´UTR Reporter vector
LightSwitch™ 3´UTR
Reporter Vector.

LightSwitch™ lncRNA Promoter Reporter Collection

While long noncoding RNAs (lncRNAs) have been shown to play important roles in the regulation of both transcription and chromatin organization, the mechanisms by which their own expression is regulated seem to differ from those of protein-coding genes. The LightSwitch lncRNA Promoter Reporter Collection was designed to make it fast & easy to study the regulation of lncRNA promoters.

LightSwitch Promoter Reporter vector
LightSwitch™ Promoter
Reporter Vector.

LightSwitch™ Validated
Pathway Collections

The Validated Pathway Collections are subsets of our LightSwitch Promoter constructs that were selected based on sequence motif analysis and published information, followed by in-house testing using our LightSwitch Luciferase Assay. All members have been experimentally validated to show significant activation or repression in response to pathway-specific induction conditions, so they are ideal when studying pathways such as inflammation, hypoxia, DNA damage, heat shock, etc.

LightSwitch Validated Pathway Collections
LightSwitch™ Validated
Pathway Collections.

Synthetic Response Elements &
Synthetic miRNA Targets

Because they have been engineered for increased sensitivity, the use of synthetic regulatory elements can improve some regulation experiments greatly. The stronger responses provided by our optimized LightSwitch Synthetic Response Elements and LightSwitch Synthetic miRNA Target Reporters can yield more meaningful data than that from native, endogenous promoter and 3´UTR sequences.

LightSwitch Synthetic Enhancers
Synthetic Enhancers &
miRNA Targets.

LightSwitch™ Custom Services

In addition to the many different products that make up the LightSwitch Luciferase Assay System, we offer a wide variety of custom services, including:

LightSwitch Custom Services
Our Custom Services Lab Can
Accelerate Your Research.

LightSwitch™ Pathway Reporter
Stable Cell Lines

Each LightSwitch Pathway Reporter Stable Cell Line contains a stably integrated, human regulatory element driving the expression of our optimized RenSP luciferase gene. This makes it far easier to measure the activity of specific pathways in response to various treatments and growth conditions. Each cell line has been validated using a variety of different plate-based formats, making them particularly useful for high-throughput screening experiments.

LightSwitch Pathway Reporter Stable Cell Lines
LightSwitch™ Pathway Reporter
Stable Cell Lines.

Transfection Reagents

After selecting a LightSwitch reporter construct, you must get it into your cell line. For plasmid transfections and co-transfections, we offer FuGENE® HD Transfection Reagent because of its consistent, high-quality performance across a wide variety of cell lines. For co-transfecting plasmids and miRNAs, we offer DharmaFECT® Duo Transfection Reagent, which provides more efficient co-transfection of those two distinct types of molecules. We also offer the LightSwitch Transfection Optimization Kit to quickly determine the conditions needed for high-efficiency transfections of your specific cell lines.

Mammalian cell transfection
Mammalian cell transfection.

LightSwitch Assay Reagents

After transfection, the amount of luciferase produced is measured using our LightSwitch Luciferase Assay Kit. Because RenSP catalyzes oxidation of its substrate in a reaction that produces light, gene expression can be quantified using a luminometer. The novel, proprietary substrate and optimized lysis buffer included in the LightSwitch Luciferase Assay Kit were formulated to provide high sensitivity over a broad dynamic range when used with our engineered RenSP. For those experiments that utilize co-transfection for normalization, we offer our LightSwitch Dual Assay Kit.

LightSwitch Luciferase Assay Kits
Light produced by luciferase.

LightSwitch Controls & Vectors

To help ensure you get the best results possible, we have developed panels of LightSwitch Promoter Control Vectors and LightSwitch 3´UTR Control Vectors. These positive & negative control constructs make it easier to interpret your data and to troubleshoot your work should things not go as expected. In addition, we offer Empty LightSwitch Reporter Vectors, which are useful as controls and for cloning your own regulatory elements.

Positive & negative LightSwitch Promoter Control Vectors
Positive & negative promoter controls.