- What antibody do you use for co-staining?
MAIT cells are known to express CD3, CD69, and CD161 on the surface. Staining with anti-CD3 antibody was used in our data collection process.
- Are there any markers other than MR1 tetramer to detect MAIT cells?
It has been reported that human MAIT cells express TCR Vα7.2. However, not all cells expressing TCR Vα7.2 are MAIT cells.
- How long can the prepared ligand be stored before loading onto the MR1 Tetramer?
Vitamin B2 metabolic intermediates are unstable in aqueous solutions. It is therefore recommended to prepare ligand solutions immediately before loading onto the tetramer. We checked tetramer staining using the MR1 Tetramer stored for 7 months after ligand loading, and the reactivity is reduced to less than a half comparing with the Tetramer whose ligand is loaded just before the assay.
- How stable is the MR1 tetramer?
We have confirmed a shelf life of up to 12 months. It is recommended that ligands are loaded just before use, and are subsequently analyzed as soon as possible.
- What is the expected positive rate of MR1 Tetramer?
When using a Tetramer staining enhancer, the positive rate is expected to be around only a few percent. However, variations may occur between individuals, and it could also happen that positive cells cannot be detected in some of the samples. Conversely, in some cases, a sufficient positive rate can be obtained without using an enhancer. In addition, the degree of non-specific background staining tends to vary greatly depending on the samples. When considering cell staining, we recommend that you use samples from as many donors as possible.