CD33 Monoclonal Antibody (WM-53 (WM53)), Brilliant Violet™ 605, eBioscience™

Product Specific Information

Description
The WM-53 monoclonal antibody reacts with human CD33, also known as GP67 and P67, a 67 kDa type I transmembrane glycoprotein that is a member of the Siglec (sialic acid-binding Ig superfamily lectin) family. It is highly specific to the hematopoietic compartment and is expressed on monocytes, activated T cells, granulocytes, myeloid progenitors, and mast cells.

Applications Tested
This WM-53 (WM53) antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (0.25 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.

Blocking Buffers
When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) or Brilliant Stain Buffer (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.

Light sensitivity
This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation
• Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) 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.
• Our internal testing suggests that Brilliant Violet™ 605 (BV605) is not compatible with methanol-based fixation.

Excitation: 407 nm; Emission: 605 nm; Laser: Violet Laser.

BRILLIANT ULTRA VIOLET™ is a trademark or registered trademark of Becton, Dickinson and Company or its affiliates, and is used under license. Powered by Sirigen™.

Target Information

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.