Single Cell Closeness Ligation Assay Service
Single cell closeness ligation assay is a method for analyzing both mRNA and protein simultaneously at single cell resolution. The final output can be measured using qPCR, fluorescently, or through DNA sequencing.
Introduction of Single Cell Closeness Ligation Assay
The Single Cell Closeness Ligation Assay (SCCLA) is a revolutionary technology that allows for the in situ detection of endogenous proteins, protein modifications, and protein interactions with high specificity and sensitivity. This method enables the direct observation of individual endogenous protein complexes in situ, providing a powerful tool for the study of protein-protein interactions at the single-cell level. SCCLA is capable of detecting and localizing protein targets with single-molecule resolution in unmodified cells and tissues, providing a detailed understanding of protein interactions and their role in cellular function.
Fig.1 Diagram of single cell closeness ligation assay. (Vistain, 2021)
Advantages of Single Cell Closeness Ligation Assay
SCCLA offers several advantages over traditional methods for studying protein interactions. Firstly, it provides high specificity and sensitivity, allowing for the detection of individual protein complexes in their natural cellular environment. This is achieved through the use of two primary antibodies raised in different species to detect two unique protein targets. A pair of oligonucleotide-labeled secondary antibodies then bind to the primary antibodies, forming a closed, circle DNA template that is required for rolling-circle amplification. This results in an amplified signal that is still tethered to the probe, allowing for the precise localization of the signal.
Secondly, SCCLA offers versatility in its application. It can be performed on a variety of sample types, including adherent cells, cytospin preparations, and tissue sections on glass slides. It can also be used in conjunction with various detection methods, such as immunofluorescence or immunohistochemistry. Furthermore, SCCLA can be performed on blood or suspension cells for detection by flow cytometry and can be used in multi-well plates for high-throughput analysis.
Creative Biolabs' Single Cell Closeness Ligation Assay
At Creative Biolabs, we offer a comprehensive Single Cell Closeness Ligation Assay service that is tailored to your specific research needs. Our team of experts has extensive experience in the field of protein-protein interactions and is equipped with the latest technology to provide accurate and reliable results.
Our SCCLA service includes the provision of all necessary reagents, the performance of the assay, and the analysis of the results. We also offer consultation and support throughout the process, ensuring that you receive the most out of your research.
For more information, please contact us.
1. What is the sensitivity of the Single Cell Closeness Ligation Assay?
The SCCLA is highly sensitive and can detect individual protein complexes in their natural cellular environment. This is achieved through the use of a rolling-circle amplification process, which results in an amplified signal that is still tethered to the probe, allowing for the precise localization of the signal.
2. Can the Single Cell Closeness Ligation Assay be used on any type of cell or tissue sample?
Yes, the SCCLA can be performed on a variety of sample types, including adherent cells and tissue sections on glass slides. It can also be used in conjunction with various detection methods, such as immunofluorescence or immunohistochemistry.
3. How does Creative Biolabs ensure the accuracy and reliability of the Single Cell Closeness Ligation Assay results?
At Creative Biolabs, we have a team of experts with extensive experience in the field of protein-protein interactions. We use the latest technology and adhere to strict quality control standards to ensure the accuracy and reliability of our results. We also offer consultation and support throughout the process, ensuring that you receive the most out of your research.
4. Can the Single Cell Closeness Ligation Assay be used for high-throughput analysis?
Yes, the SCCLA can be performed on blood or suspension cells for detection by flow cytometry and can be used in multi-well plates, such as 96- or 384-well plates, with a high-content screening imager for high-throughput analysis.
5. What are the applications of the Single Cell Closeness Ligation Assay?
The SCCLA is a powerful tool for studying protein-protein interactions and their role in cellular function. It can be used for pathway analysis, drug screen kinetics, IC50 determination, and target validation. It is also a valuable tool for the study of diseases associated with protein interactions, such as cancer.
Reference
- Vistain, L.F.; Tay, S. Single-cell proteomics. Trends in Biochemical Sciences. 2021, 46(8): 661-672.
