High-quality and reliable fluorescent antibodies are the guarantee for the successful completion of flow assays and their related data analysis, especially for multicolor flow assays based on fluorescence compensation and requiring high resolution. How to choose high quality fluorescent antibodies for flow cytometry? Here are some key factors.
Three key factors
1. Common clone number
For a specific cell surface antigen CD family protein, there are usually multiple monoclonal antibodies with different clone numbers. The more common the clone number used in a flow cytometry experiment, the greater the chance of success.
2. High Staining Index
A high staining index facilitates the separation of positive and negative cell populations, especially if the expression of the target protein is low.
3. Low background binding
Although the background can usually be reduced by using an isotype control or Fc Blocking Reagent, poor coupling of fluorescent dyes to antibodies can significantly exacerbate background binding. Lowering the background makes it easier to define positive and negative cells.
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