Frequently Asked Questions

You Asked, We Answered

 

Have more questions? Contact Tech Support!

We're here to help you with any task or project you may have! Feel free to contact us with any of your inquiries!

arrow&v

Thanks for submitting!

 

What differentiates GenuIN Biotech from other competitors in the antibody market?

In Western blotting, if a band at the expected size can be detected by an antibody in wild-type (WT) cell lysates but disappears or significantly diminishes in gene-silenced cell lysates, the antibody is considered to be specific. Similarly, if an antibody can stain the WT cells (as indicated by strong fluorescent signals), but the signals are either lost or significantly reduced in gene-silenced cells, this antibody is deemed specific in immunofluorescence (IF) or flow cytometry (F) applications. Based on our criteria, an average of 30% antibodies tested are specific for use in Western blotting. When it comes to immunocytochemistry and immunofluorescence (IC-IF) or flow cytometry (F), only 20% of antibodies are specific.

 

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 instance, we developed 8 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.

What percentage of antibodies in the market have passed your rigorous testing?

In Western blotting, if a band at the expected size can be detected by an antibody in wild-type (WT) cell lysates but disappears or significantly diminishes in gene-silenced cell lysates, the antibody is considered to be specific. Similarly, if an antibody can stain the WT cells (as indicated by strong fluorescent signals), but the signals are either lost or significantly reduced in gene-silenced cells, this antibody is deemed specific in immunofluorescence (IF) or flow cytometry (F) applications. Based on our criteria, an average of 30% antibodies tested are specific for use in Western blotting. When it comes to immunocytochemistry and immunofluorescence (IC-IF) or flow cytometry (F), only 20% of antibodies are specific.

Since the beginning of the COVID-19 pandemic, a large number of monoclonal and polyclonal antibodies against SARS-CoV-2 N and S proteins have entered the antibody market, most of which have been validated with recombinant proteins. There is no doubt that these antibodies react with their respective antigens, however, whether they work with the actual virus or human samples still need to be tested. In our experience, only 15% of antibodies that reacted with their respective antigens functioned well with SARS-CoV-2 and human specimens.

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.

Do we have to use your reagents together with your antibodies in our experiments?

At certain critical points, we strongly recommend using our reagents. Our experiences show that certain proteins in the whole cell lysate are at picogram and even femtogram levels. Detecting these proteins in Western blotting using regular ECL reagents is a seemingly impossible task. In this case, our custom PiQ (Cat#636) or FeQ (Cat#226) ECL reagents can detect picogram- and femtogram-level proteins, respectively. Second, we recommend you always use our FadeStop fluorescence mounting medium (Cat#270) or FadeStop fluorescence mounting medium with DAPI (Cat#272, to counterstain nuclei) in immunofluorescent staining. These antifade fluorescence mounting media have passed our testing and can preserve fluorescence after two months when stored at -20°C or after as many as twelve laser scans using a confocal microscope. Finally, we recommend using our IntactProteinTM Cell Lysis Kit (Cat#415) to extract whole cell lysates from cultured mammalian cells. This kit is particularly formulated to maintain the integrity as well as the signal moiety (such as phosphorylation) of all sizes of proteins. Since this kit avoids sonication, it protects proteins, particularly large-sized proteins (> 100 kDa), from fragmentation.        

Why do you supply more than one monoclonal antibody for certain proteins?

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 samples can change the conformation of the epitope recognized by the antibody; therefore, it is not surprising that an antibody functions well in Western blotting, but does not detect the antigen in immunohistochemical staining; 3) For multiple staining on the same samples such as tissues or cells, a combination of  mouse and rabbit antibodies is preferrable; 4) For some sandwich ELISA, you need two antibody clones from the same or two different species.  

Can I use an antibody for applications other than those stated in your data sheet?

Yes, but we cannot guarantee that it will work. Our antibodies are only tested for use in Western blotting (WB), immunocytochemistry and immunofluorescence (IC-IF), and flow cytometry (F) applications. However, it is highly probable that these validated antibodies can be used in other applications.