How are your antibodies against SARS-CoV-2 distinguished from other vendors?
Most SARS-CoV-2 monoclonal antibodies are developed using recombinant N or S proteins. These antibodies will react with the recombinant proteins; however, they may not react with true viruses for a number of reasons. For instance, we developed 8 rabbit monoclonal anti-S antibodies using recombinant S proteins, only 1 of which worked with SARS-CoV-2 and human specimens. We have validated our antibodies using real coronavirus and human nasopharyngeal swabs in different applications so that you can be sure that these antibodies work without trying and error.
Why do you supply more than one monoclonal antibody for N or S protein?
Monoclonal antibodies, especially recombinant antibodies, are an ideal tool for investigating SARS-CoV-2 infection because they have minimal batch-to-batch variation and produce consistent and reproducible results. We provide multiple clones for a given protein for the following reasons: 1) Each monoclonal antibody recognizes a specific epitope on a protein, you can select the one that best suits your purposes; 2) Different methods of processing viruses and/or specimens can change the conformation of the epitope recognized by the antibody; therefore, it is not surprising that an antibody function well in Western blotting, but does not detect the virus in immunohistochemical staining; 3) For multiple staining on the same samples such as tissue or cells infected with SARS-CoV-2, a combination of mouse and rabbit antibodies is preferable; 4) For some sandwich ELISA, you need two antibody clones from the same or two different species.
What are the benefits of using your validated shRNA lentiviruses?
We provide human shRNA lentiviral particles to facilitate your mechanism research. Using our proprietary lentiviral platform, we have designed and tested the specificity of these viruses using Western blotting, immunocytochemical staining, or flow cytometry. What you need to do is to put the viral medium onto your cultured cells and select the cells to be 100% pure with an antibiotic known as puromycin. Additionally, you will obtain a stable cell line with your gene/protein of interest knocked down. The benefit for you is that we validate the specificity of these viruses so that you don’t need to do it all out of scratches, saving your resources, and above all, your time.
Can you give me an example of how lentiviral shRNA can be used in the study of SARS-CoV-2 infection?
Let’s say I plan to investigate whether STAT3 is involved in the JAK-STAT signaling pathway triggered by SARS-CoV-2 infection, I would apply the validated STAT3 shRNA lentiviruses onto the cells of interest (Cat# V7713). After confirming that STAT3 has been silenced, I can use this cell line to perform a series of studies. The same principle applies if I want to study multiple pathways, I can knock down several proteins using their specific lentiviral shRNAs.