ALDEFLUOR Stem Cell Identification Kit

Based on aldehyde dehydrogenase (ALDH) enzymatic activity, this fluorescent assay enables identification and isolation of stem and progenitor cell populations by flow cytometry. The kit, supplied by STEMCELL Technologies, contains a lyophilized BODIPY-aminoacetaldehyde (BAAA) substrate, proprietary assay buffer, and a DEAB (diethylaminobenzaldehyde) vial used as the negative-control inhibitor.

In the workflow, BAAA freely diffuses into cells and is converted intracellularly by ALDH into a charged fluorescent product (BAA) that is retained inside the cell. Cells exhibiting high ALDH activity, termed ALDH-bright, are detected with a 488 nm blue laser and emission captured in the FITC/green channel near 530 nm. The parallel DEAB control tube blocks ALDH activity, establishing a fluorescence baseline. Approximately 100 tests can be performed per kit, with reagents handled in single-use aliquots.

• Isolation of human hematopoietic stem and progenitor cells from umbilical cord blood, bone marrow, and mobilized peripheral blood
• Identification of cancer stem cell populations in leukemia and solid tumors including breast, lung, and colon
• Validation of ALDH-bright fractions by colony-forming assays or xenograft transplantation experiments

Protocol steps involve 30 to 60 minute incubations at 37°C followed by flow cytometry or fluorescence-activated cell sorting. Storage requirements include the BAAA substrate vial kept frozen at -20°C and protected from light, while the assay buffer is refrigerated. Lyophilized components remain stable until the printed expiration when stored properly. For research use only.