background:

Reporter gene detection is an important means of studying gene expression regulation and has been widely used in the fields of molecular biology and cell biology research. Reporter gene detection is a method that couples transcriptional regulation and reporter gene (such as luciferase) expression, and determines the biological activity of cell stimulants (such as small molecule compounds, recombinant proteins) by measuring the activity of reporter gene expression products (such as luciferase). The 2015 edition of the Chinese Pharmacopoeia has listed the reporter gene detection method as the second method of interferon activity determination (General Rule 3523). At present, the reporter gene detection method has played a vital role in the biological activity detection and Fc effector function research of related monoclonal antibody drugs such as biological products PD1/PD-L1, IL6/IL6R. In the past, cytokines such as EPO, GH, FSH, TSH, etc., which could only be studied by in vivo animal biological activity, are also gradually adopting in vitro reporter gene detection methods to replace the original in vivo animal method as the key indicator for product batch release. Transcriptional regulation combined with reporter gene expression is often used in a wide range of physiological studies, such as analyzing receptor function by quantifying the effect of specific receptor response elements on gene expression, or for studying signal transduction, transcription factors, protein-protein interactions, and viral infection and transmission. Events downstream of transcription, such as mRNA processing and protein folding, can also be analyzed. Therefore, luciferase is widely used in the biotechnology and pharmaceutical industries.

Firefly luciferase is a protein with a molecular weight of 61kDa. It has no background fluorescence in host cells , does not require post-translational modification, and is fast, reliable and convenient to detect. It is an ideal choice for detection using the firefly luciferase reporter gene system. This kit uses D-luciferin as a substrate. In the presence of ATP, magnesium ions and oxygen, it can be catalyzed by firefly luciferase to undergo oxidative decarboxylation to produce activated oxyluciferin and bioluminescence ( Figure 1). Bioluminescence is proportional to the promoter activity of luciferase, which indirectly reflects the biological activity of the cell stimulant.

Features:

The signal half-life of this product is 30 minutes at room temperature (22-25°C); Bio- One ^TM One-Step Firefly Luciferase Assay Kit has the characteristics of easy operation, stable reagents, ultra-high light output, and high sensitivity.

1. Convenient: Simply mix the freeze-dried powder substrate with the buffer solution and it is ready for use. The sample detection steps are simple;

2. Fast: luciferase detection is completed within 2-15 minutes;

3. Ultra-high light output: Stronger light output, can produce up to 10 times higher light intensity than other luciferase detection reagents;

4. High sensitivity: Able to detect as low as 10 ^-20 mol of luciferase.

Specifications:

Storage conditions:

Transported in ice packs. After receiving the test kit, it can be stored at -30 ~ -15℃ for a long time. To ensure the excellent performance of the mixed reagents, it is recommended to store the unused reagents at -70℃.

application:

Bio- One ^TM One-Step Firefly Luciferase Assay Kit is mainly suitable for firefly luciferase reporter gene systems that require ultra-high light output signals.

Note:

We also provide a series of ADCC and ADCP effector cell lines. Please contact techserv@rhinobio.com for more information.

Frequently asked questions

Q1: No signal or low signal strength

A1: This problem may occur for the following reasons: low activity or failure of luciferase assay reagent, low expression of luciferase , etc. The corresponding solutions or precautions are as follows: