Can I use the Pro-Detect assay kits for rapidly isotyping antibodies?
The Pro-Detect Rapid Antibody Isotyping Kit is a lateral-flow assay with high sensitivity for rapid determination of the immunoglobulin class, subclass, and light chain type of monoclonal antibodies. Two different formats of rapid isotyping kits are available. These include dipstick format and cassette-based format. One can use these kits to identify the isotype of a mouse, rat, or human antibody. Make sure to follow the recommended dilution guidelines.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
I am using a Pro-Detect assay kit and cannot differentiate banding pattern between control lysates and lysates that are actually expressing proteins of interest. What is the problem?
1) It is likely that there could be a substance interfering with the assay. Check the compatibility table on this page (https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-assays-analysis/pro-detect-rapid-assays.html), under Detergent and salt compatibility limits.
2) The expression level of the actual tagged protein is below the detectable range of the assay.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
Using Pro-Detect assay kits, can I detect the protein of interest in a mixture of proteins such as a crude lysate?
Yes, first, make sure that the protein of interest is tagged and that we offer a Pro-Detect assay kit that can detect that tag. Secondly, make sure that you follow the guidelines to detect proteins in crude lysate and if necessary increase or decrease the lysate concentration to bring the protein of interest within the detectable range of the assay.
Note that the guidelines for the recommended detection ranges reflect the concentration of the pure tagged protein in a given sample mixture.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
How do I interpret the gradual disappearance of four lines in the Pro-Detect competition assay kits?
As the concentration of the tag in the protein sample increases, the number of lines will decrease until they all disappear. The concentration of the tagged proteins is inversely related to the number of lines appearing on the strip.
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Are the results of Pro-Detect assay kits comparable to western blotting?
Yes. Western blotting is a widely used and accepted tool to identify a specific protein in a complex mixture. However, it is a time-consuming and labor-intensive process, with many preparative steps and time intervals to manage. The Pro-Detect rapid assays are able to provide comparable accuracy but with much shorter processing time (10-15 min) and less hands-on steps. For further information, please see the data shown on this page (https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-assays-analysis/pro-detect-rapid-assays.html) under Results are comparable to western blotting.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.