Background / objective:
Hematopoietic neoplasms lead to disease states that involve abnormal proliferation of blood cells. Ki-67 and carboxyfluorescein succinimidyl ester (CFSE) is assays used to examine the proliferation status of cells but affect cell viability. In this study, we use lectins to bind to surfaces of proliferating cells with different phenotypes while preserving cell viability.
Materials and methods:
The Friend mouse lymphocyte leukemia cell line F5-5.F1 was stained using biotin-conjugated lectins from Canavalia ensiformis (ConA), Dolichos biflorus (DBA), Erythrina crista Galli (ECA), Lens culinaris (LCA), Phaseolus vulgaris (PHA-E4), Arachis hypogaea (PNA), Ulex europaeus (UEA) and Triticum Vulgaris (WGA) and classified by fluorescence-activated cell sorting. Morphology, gene expression, and proliferation assays were performed in sorted cells.
DBA, LCA, and PHA-E4 probing cells sorted according to surface phenotype. Gene expression analysis showed that the myelocytomatosis oncogene (Myc), cyclin D1 (Ccnd1), and cyclin D2 (Ccnd2) were more expressed in the DBA (high) fraction than the DBA (Int) and DBA (Neg) fractions. Ki-67 expression and the MTS assay correlated with the DBA binding pattern, with DBA (High) reflecting the highest proliferative tendency.
DBA labeling allows the selection of proliferating cells by flow cytometry.
Lectin; cell proliferation; flow cytometry; leukemia.
Unit size: 5 mg
Applications: Immunohistochemistry / Immunocytochemistry, Immunofluorescence, Transfer Applications, Elispot, ELISA, Glycobiology
Recommended Use: For most applications, we recommend a freshly prepared 5-20 µg / ml working solution in the above buffer.
Recommended storage: 2-8 ° C; Store frozen for long-term storage
Solution: 10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.08% sodium azide, 0.1 mM CaCl2.
Concentration: 5 mg active conjugate / ml
Sugar specificity: N-acetylgalactosamine