ANTI-HUMAN CD33 (WM53) COLORWHEEL DYE-READY MAB

General description

ColorWheel^® technology is a novel and proprietary method of creating your own antibody and dye combinations for use in flow cytometry. By incubating any ColorWheel^® antibody with any ColorWheel^® dye, researchers can quickly and simply produce primary conjugated antibodies for use in single color or multicolor flow cytometry analysis. Each ColorWheel^® antibody and dye is lyophilized for long long-term storage, allowing for simplicity and flexibility without compromise. ColorWheel^® technology requires both ColorWheel^® antibody and ColorWheel^® dye for Flow Cytometry application.

We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This product is Preservative-free, lyophilized product for enhanced stability and allow for ambient shipping and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.

Specificity

Clone WM53 is a mouse monoclonal antibody that detects human Myeloid cell surface antigen CD33. It targets an epitope within the V-Ig-like domain of isoform 1 (CD33M).

Immunogen

Acute Myeloid Leukemia (AML) cells.

Application

Quality Control Testing:

Evaluated by Flow Cytometry in Human peripheral blood mononuclear cells (PBMC).

Flow Cytometry Analysis: Staining of one million Human peripheral blood mononuclear cells (PBMC) was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1004, Anti-CD33 Antibody (WM53) ColorWheel^® Mouse mAb and Cat. No. CWD-PE ColorWheel^® activated Phycoerythrin (PE) Dye or an equivalent amount of PE-conjugated Mouse IgG1 isotype control.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Compatibility

Requires ColorWheel^® Dye (Sold Separately)

Target description

Myeloid cell surface antigen CD33 (UniProt: P20138; Sialic acid-binding Ig-like lectin 3, Siglec-3, gp67, CD33) is encoded by the CD33 (also known as SIGLEC3) gene (Gene ID: 945) in human. CD33 is a disulfide-linked homodimeric, single-pass type I membrane glycoprotein that is synthesized with a signal peptide (aa 1-17), which is subsequently cleaved off to generate the mature form that contains an extracellular domain (aa 18-259), a transmembrane domain (aa 260-282), and a cytoplasmic domain (aa 283-364). In its extracellular region it contains one Ig-like V-type domain (aa 19-135) and a Ig-Like C2 type domain (aa 145-228). In its cytoplasmic domain it has two ITIM motifs (aa 338-343 and 356-361) that are involved in modulation of cellular responses. These motifs can be phosphorylated by Src-like kinases and phosphorylated ITIM motifs can bind to SH2 domains of several SH2-containing phosphatases. Phosphorylation of CD33 by LCK on tyrosine 340 is involved in its binding to PTPN6 and PTPN11. CD33 is mainly expressed in monocytic/myeloid lineage cells. Its expression is also observed on microglial cells in the brain. It plays a role in mediating cell-cell interactions and in maintaining immune cells in a resting state. It is present in a vast majority of subjects with Acute Myeloid Leukemia (AML), but is absent from pluripotent hematopoietic stem cells. Clone WM53 is reported to specifically detect CD33M isoform. It recognizes an epitope located in the Ig-like V type domain, which is only expressed in CD33M. (Ref.: Walter, RB., et al. (2012). Blood. 119(26); 6198-6208; Pérez-Oliva, AB., et al. (2011). Glycobiology. 21(6); 757-770; Taylor, VC., et al. (1999). J. Biol. Chem. 274(17); 11505-11512).

Physical form

Lyophilized from PBS containing D-Mannitol and Sucrose, normal appearance is a dried pellet. Reconstituted antibody solution is stable and functional as assessed by functional testing. Contains no biocide or preservatives, such as azide.

Reconstitution

1.0 mg/mL after reconstitution at 25 μL in PBS. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommend storage of lyophilized product at 2-8°C. Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial. Reconstitute each vial by adding 25 μL of PBS. Protect from light.

Legal Information

ColorWheel is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.