Cignal Reporter Controls

Cignal Reporter Assayを用いた際のコントロール実験

S_2798_ADNA_CignalReporter_s
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Cignal Reporter Controls

Cat. No. / ID:  336881

Cignal Reporter Control
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¥46,500
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Cignal Reporter Controlsは分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。
Configure at GeneGlobe
Find or custom design the right target-specific assays and panels to research your biological targets of interest.

Features

  • トランスフェクション条件の至適化に最適
  • 処理効果の特異性を確立
  • 即トランスフェクション可能
  • 高い感度と特異性

Product Details

Cignal Reporter Assayは、 ダウンストリームの転写因子活性を測定することにより、シグナルパスウェイの活性を迅速かつ高感度で定量的に測定します。Cignal Reporter Assayを用いて実験を実施する際に、適切なコントロールが実験計画の重要な要素になります。これにより、レポーターアッセイ結果を正確に分析することができ、観察された応答の特異性の確信が得られます。

Principle

Cignal Reporter Controlはトランスフェクションが即可能な調製済みコンストラクトです。ポジティブおよびネガティブコントロールは、デュアルルシフェラーゼおよびGFPレポーター用が入手可能です。

次のコントロールが入手可能です。

入手可能なコントロール
コントロール 概要
Positive Control Assay(GFP) GFP遺伝子を恒常的に発現。GFP(green fluorescent protein)を用いて導入効率の測定や導入条件の至適化が容易
Negative Control Assay(GFP) 非誘導性、最小プロモーターで発現するGFP遺伝子。バックグランドのGFP蛍光を測定し、処理効果の特異性を確立
Positive Control Assay(luc) 恒常的発現GFP、恒常的発現ホタルルシフェラーゼ、恒常的発現ウミシイタケルシフェラーゼのコンストラクトが40:1:1で混和。導入効率を測定し、ホタルルシフェラーゼアッセイのポジティブコントロールとして機能
Negative Control Assay(luc) 非誘導性ホタルルシフェラーゼコンストラクトと恒常的発現ウミシイタケルシフェラーゼコンストラクトが40:1で混和。処理効果の特異性を確立

Procedure

Cignal Lenti Reporter Control Assayはウイルス力価を測定済みの、即導入が可能なレンチウイルス粒子で、導入条件の至適化に使用します。標準プロトコールを用いてCignal Lenti Reporter Control Assayを選択細胞に導入し、レポーター導入細胞と同様に、コントロールを導入した細胞を処理します。

Applications

Cignal Reporter Assayを用いて実験を行なう際に、Cignal Reporter Controlは実験計画の重要なエレメントになります。以下のような多様なツールとして機能します:  

  • 細胞システムへの導入条件の至適化  
  • トランスフェクション効率の定量  
  • 処理効果の特異性を確立 
  • バックグランドのGFP蛍光あるいはルシフェラーゼ活性の測定  
  • 標準化のための内部コントロールとして機能

Supporting data and figures

Resources

Transfection Protocols (2)
Search for transfection data by nucleic acid, cell line, and transfection reagent. Our database contains data from researchers like yourself who have shared their experimental results with us.
Transfection protocols for specific cell types and plate formats that save you the time and effort of adapting existing protocols to fit your requirements. Simply select the cell type, nucleic acid, and culture format to receive a QIAGEN transfection protocol to print out or download in convenient PDF format.
MSDS (1)
Download Safety Data Sheets for QIAGEN product components.
キットハンドブック (1)
For cell-based pathway activity assays
パンフレット (1)
For cell-based analysis of pathway signaling activity
Safety Data Sheets (1)
Certificates of Analysis (1)
Brochures & Guides (1)
For cell-based analysis of pathway signaling activity
Kit Handbooks (1)
For cell-based pathway activity assays

FAQ

What transfection reagents are compatible with the Cignal Reporter Assays?
Any transfection reagent that yields 25-30% transfection efficiency in the cell line of interest is suitable for use with DNA-based Cignal Reporter Assays.
FAQ ID -2764
Can users insert custom transcriptional regulatory elements or promoters into the Cignal Reporter Assays?
No, The Cignal Reporter Assays do not currently support this functionality.
FAQ ID -2765
Can I transform the Cignal Reporter Assays?
No, DNA-based Cignal Reporter Assays are not designed for bacterial transformations.
FAQ ID -2762
The pathway reporter luciferase activity values are greater than the positive control, is there a problem?
No, the positive control only serves as a constitutively-active reporter enabling the user to assess if the transfection/transduction was successful, and should not be used for direct comparison to the pathway reporters.
FAQ ID -2766
What is MOI?
MOI is an abbreviation for Multiplicity of Infection or the number of viral particles exposed to a cell.
FAQ ID -2768
What MOI should I use for my cells?
This has to be empirically determined by the user. By using a positive control, Cignal Lenti Reporter Control,  the user can set up parallel transductions with increasing amounts of virus to identify an MOI that yields a robust signal.
FAQ ID -2770
The pathway reporter luciferase activity values are less than the negative control, is there a problem?
No, The negative control only serves as a non-inducible reporter, but should not be used for direct comparison to the pathway reporters.
FAQ ID -2767
Can I make stable cell lines with the Cignal Reporter Assays?

No, the DNA-based Cignal Reporter Assays are not designed to generate stable pathway sensor cell lines. However, the Cignal Lenti Reporter Assays can be used to generate stable pathway sensor cell lines.

FAQ ID -2763
What is the concentration of SureENTRY Transduction Reagent?
The concentration of SureENTRY Transduction Reagent is 10 mg/ml.
FAQ ID - 3708
How do I choose between the DNA-based Cignal Reporter Assays and the Cignal Lenti Reporter Assays?
The DNA-based Reporters are designed for cells that are amenable to transfection. QIAGEN recommends a minimum transfection efficiency of 25 – 30%. If the cells are refractory to transfection then the Cignal Lenti reporters should be selected.
FAQ ID -2761
How many experiments can I do with the Cignal Lenti Reporter Assays?
This depends on the Multiplicity of Infection (MOI) required for a specific cell line. The lower the MOI the more cells that can be transduced. Therefore, to predict how much Cignal Lenti Reporter is required an MOI must be determined.
FAQ ID -2769