With the Premo™ Autophagy TB/GFP TR-FRET LC3B Expression Kit you get:
- An objective method to quantify autophagy
- The ability to assay autophagy in disease-relevant cell lines
- A scalable, plate-based format
- Correlation with historical data
- Assay development in 2-3 weeks
The Premo™ TR-FRET LC3B Expression kit gives you the ability to not only see autophagy, but to quantify it. The measurement of autophagy has traditionally been performed using Western blot, fluorescence microscopy, and high content analysis. These methods are typically visual readouts, which are subject to interpretation. Subjective results make it difficult to compare experimental results. By obtaining quantitative data with the Premo™ TR-FRET LC3B assay, conditions and experiments can be easily and objectively compared.
Obtain Disease-Relevant Results
Much about the autophagic process is still not well understood, including its role in cancer and neurodegeneration. Researchers need to assess the autophagic response in disease-relevant cell models in order to extrapolate in vitro data to an in vivo response. Because it utilizes a BacMam 2.0 reagent encoded with a GFP-tagged LC3B protein, the Premo™ TR-FRET LC3B Expression Kit enables measurement of autophagy in disease-relevant cell lines without the need to make stable cell lines. As a result, assay development is 2-3 weeks rather than 2-3 months.
Generate More Data in Less Time
Traditional autophagy assay methods such as Western blot, fluorescence microscopy, and flow cytometry are high touch and low throughput assay methods, making simultaneous assessment of multiple conditions very difficult. The Premo™ TR-FRET LC3B Assay is a 96- or 384-well assay, enabling straightforward experimental scaling so you can gather more data. Traditional assay methods also require more than 5 sample manipulation steps. The Premo™ TR-FRET LC3B assay workflow requires 3 addition-only steps to obtain quantitative data (see Figure), giving you back your time.
Produce Results Consistent with Historical Data
The Premo™ TR-FRET LC3B assay has been benchmarked against known modulators of autophagy and against traditional assay methods. When the effects of autophagy modulators Wortmannin, PI-103, and Tamoxifen, were assayed with the Premo™ Autophagy Terbium/GFP TR-FRET LC3B Assay, results were consentient with known pharmacology as compared to traditional methods (see FAQ guide for data). You can be confident that the results obtained with the Premo™ TR-FRET LC3B assay will correlate to historical data.
The Premo™ Autophagy TB/GFP TR-FRET LC3B kit contains :
- BacMam 2.0 reagent encoding a GFP-tagged LC3B protein
- Terbium-labeled LC3B antibody
- Instrument control to ensure proper plate reader set-up
- 6x lysis buffer
- Chloroquine positive control