The flow cytometry market is filled with an abundance of products for mouse and human samples. But what if your studies use different species? Fortunately, many antibodies for standard cell markers can work on multiple species, and more species-specific reagents are becoming available.
Flow cytometry is an ideal way to do immune profiling analysis to measure the presence and relative proportions of cellular components of the immune system. Immunophenotyping protocols exist for many species now, so check out these tips to help you explore your options.
Do the Right Search
Many commercially available reagents are sold for use on rat, mouse, human, and non-human primate samples. But remember to keep your flow cytometry lingo straight. A hamster anti-mouse antibody is an antibody that recognizes a marker on a mouse cell, but was made using a hamster. This distinction is also important if you need a fluorescent secondary antibody; in this case, you will need an anti-hamster antibody that can bind to the hamster-derived anti-mouse antibody.
Check Cross-reactivity
Many of the lineage markers used for immunophenotyping, like CD3, CD4, CD21, and CD14, show significant sequence and structural similarity across species. In fact, many of the commercially available antibodies developed for immunophenotyping human immune cells have been shown to detect the same immune cells in non-human primates and other mammals like dogs and pigs. Explore your commercial antibody options and check for cross reactivity with your species of interest, whether it is porcine, canine, or chicken, in the product details.
Take the Time to Test
A manufacturer may state that an antibody is cross-reactive with your species, but take the time to test and titrate potential antibodies to validate that they do recognize samples derived from your particular animal specimen.
Develop a Panel
Consider which cell types you want to characterize in your immunophenotyping panel. This may be constrained by the antibodies that you have identified to function for your particular species. At a minimum, you should aim to create a panel that distinguishes lymphocytes from myeloid cells and may further characterize lymphocyte subsets including B cells and T cells.
These basic guidelines should help you get started on basic immunophenotyping experiments in alternative species.