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Description: The HIM3-4 monoclonal antibody reacts with human CD33, a 67 kDa member of the sialoadhesion family. CD33 is expressed by myelomonocytic precursors, monocytes, mast cells, and granulocytes. Hematopoietic stem cells and lymphocytes do not express this antigen. CD33 plays a role in sialic-acid dependent cell adhesion.
Applications Reported: The HIM3-4 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This HIM3-4 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (1 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
Light sensitivity: This tandem dye is sensitive photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-822-49) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488-561 nm; Emission: 667 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
Filtration: 0.2 µm post-manufacturing filtered.
CD33 is a transmembrane protein of the sialic acid-binding immunoglobulin-like lectin (Siglec) family. It belongs to the immunoreceptor tyrosine-based inhibitory motif (ITIM)-containing molecules able of recruiting protein tyrosine phosphatases SHP-1 and SHP-2 to signal assemblies, and these ITIMs are also used for ubiquitin-mediated removal of the receptor from the cell surface. CD33 is expressed on cells of myelomonocytic lineage, binds sialic acid residues in N- and O-glycans on cell surfaces, and is a therapeutic target for acute myeloid leukemia. Further, CD33 is found on granulocyte and macrophage precursors in the bone marrow, but is not on pluripotent stem cells. CD33 is also expressed on, and is a useful marker for, peripheral monocytes. CD33 is useful for distinguishing myelogenous leukemia cells from lymphoid or erythroid leukemias. Diseases associated with CD43 dysfunction include gallbladder lymphoma and extracutaneous mastocytoma.
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