We developed this whole blood immunophenotyping panel with the aim to monitor and quantify major lymphocyte subsets (CD4+, CD8+, CD4+CD8+ αβ T cells, γδ‐T cells, B and NK cells) and monocytes in pigs. The panel involved the use of commercially available reagents, avoiding secondary antibody staining or in‐house antibody conjugations, with the aim to make the assay accessible and reproducible across laboratories. The assay is accurate, robust and represents a useful tool for immune monitoring of swine in the pharmacology and toxicology fields, or to monitor the immune status in response to vaccination and diseases.