One broadly accessible way of studying glycans is to detect them with binding reagents, much like detecting proteins with antibodies, in assays such as histology, immunofluorescence, blotting, and pull-downs. While most of the general techniques used are very similar to their antibody counterparts, glycan detection reagents come with a whole host of considerations. For instance, the most widely used glycan detection reagents are plant-derived lectins that are active only as oligomers in special buffers, have high lot-to-lot variations, and are sometimes glycosylated themselves. Our Lectenz® reagents offer a way around these issues, with each functioning as a monomer, being recombinantly expressed, and having well-defined buffer compatibility without the need of divalent metal ions. Explore all the exciting applications below and see for yourself how our SiaFindTM reagents make glycoscience simple!
Western blotting is a technique that uses binding agents, such as antibodies or lectins, to detect the presence of antigens in an immobilized sample on a membrane. For a basic overview, see the video and protocol below.
Western Blotting Protocol
Immunoprecipitation is an affinity purification technique that uses binding agents immobilized to solid matrix, such as agarose or magnetic beads, to purify antigens from solutions. The technique can also be used to purify biological complexes out of solution as long as one member of the complex contains the antigen corresponding to the binding agent. For use with our glycan detection reagents, see the protocol below.
Bovine serum IP using biotinylated SiaFindTM Pan-Specific Lectenz and streptavidin magnetic beads. The sialylated serum globulin, serpin G1, is among the glycoproteins pulled down from serum solution, as seen in the western blot.
Immunoprecipitation Protocol
Immunohistochemistry is a technique that uses binding agents, such as antibodies or lectins, to detect and stain antigens in a tissue section. While the general procedure is similar for proteins and glycans, there are distinct differences in the procedure due to the biological differences between glycans and proteins. For use with our glycan detection reagents, see the protocol below.
Immunohistochemistry Protocol
Enzyme-Linked Immunosorbent Assay (ELISA) is a plate-based technique that uses binding agents, such antibodies or lectins, to detect and quantify antigens in a sample. There are many variations in the design of the assay, such as direct, indirect, and sandwich ELISAs. To perform a basic ELISA with our glycan detection reagents, see the protocol below.
ELISA Protocol
Flow cytometry is a technique that uses lasers to measure the fluorescence of single cells or particles in solution.
Fluorescently labeled SiaFindTM (a) Pan-Specific Lectenz® or (b) α2,3-Specific Lectenz® was used to stain MDA MB231 human breast cancer cells. Overlay of untreated cells (black) and neuraminidase treated cells (red).