Here’s Everything You Should Know About Fc Block in Flow Cytometry
Like any other living thing, each cell has its own life cycle. When it comes to analyzing cells in a lab setting, flow cytometry is a widely used and comprehensive single-cell analysis method. When sorting cells or working on an experiment with flow cytometry, researchers focus on living cells. Dead cells can cause issues in the flow cytometry process and impede the results.
As you know it is very important to have good data and results, it is equally imperative to have the right controls to interpret and troubleshoot your experiment. If you want to identify whether your result is unique or artificial, or need to gate your populations accurately, with the right set of controls you can better get context and clarity.
If you find yourself wondering “What is Fc Block?” or “What is its importance during the cell separation procedure?”, this article is for you!
What is Fc Blocking?
Flow cytometry entails fluorescently labeled antibodies to bind and identify specific cellular subsets.
According to a study published by the Journal of Quantitative Cell Science, “Like cellular auto fluorescence, non-specific antibody binding introduces background noise, thereby limiting resolution and impairing the ability to identify cells with low marker expression.” The intricacies of binding rely on various factors of each antibody clone. As the Fc receptors bind with antibodies via their constant Fc domain rather than the antigen-specific Fab domain, it leads to false-positive and flawed results. To prevent this type of binding, Fc blocking reagents when added to staining protocols can ensure that only antigen-specific binding is observed.
What is the Importance of Using an FcR Blocker During Cell Separation Procedures?
During the cell separation process, it is imperative to prevent non-specific binding to ensure the purity of your cells which will, in turn, lead to optimum results. Fc receptors (FcR) on cells have a high affinity for the Fc region of antibodies. To prevent the non-specific binding of antibodies to cells, it is advisable to add an FcR blocker to your sample prior to cell separation. To prevent the nonspecific binding of the antibodies, it is recommended to add an FcR blocker to your sample before the cell separation.
While for human and mouse samples you can get cell separation kits, there isn’t any universal blocker that would work for all species. So, if you are working with a species other than a human or mouse, you will have to add a blocker appropriate to the species with which you are working.
Antibody-related protocols in long-term research projects can be exhausting and overwhelming. This anxiety is further reified by the frequent lack of long-term assurances of antibody access. If you want to get the Purified Bulk Antibody online, you can reach out to Tonbo Biosciences.
Author’s Bio- The author is a researcher, and this article is about the Fc Blocking.