This is a protocol used to perform immunophenotyping of whole blood (collected in EDTA tubes) or cells from bronchoalveolar lavage (BAL). We have successfully used this protocol for rhesus macaques, cynomolgus macaques, and African green monkeys (although antibody mix provided was titrated on AGM only). It allows the characterization of T cells (CD69/CD25 for activation; CCR7/CD45RA for memory phenotype; α/β vs γ/δ ), B cells, Monocytes, Neutrophils, Basophils, Eosinophils, an potentially NK cells (CD56 does not work for AGM).
This protocol was designed to deal with samples coming from containment labs (> CL2) at our facility. If you are using this protocol at a different facility please ensure that proper testing and approvals are in place. The protocol can be used for experiments completed entirely in CL2, simply go from step 17 directly to 31.