Description
Gemtuzumab297.rMAb is a research-grade recombinant human IgG1, κ monoclonal antibody that specifically binds the long isoform of CD33. It incorporates the same variable region sequence as the therapeutic humanized Gemtuzumab antibody—originally derived from the mouse P67.6 clone—combined with a human IgG1 constant region. To minimize Fc-mediated interactions, the conserved glycosylation site at position 297 has been replaced with alanine (N297A mutation). CD33 is a type I transmembrane glycoprotein that is a sialic acid-binding Ig-like lectin 3 (Siglec-3) of the Ig superfamily. It is expressed on myeloid progenitors, monocytes, macrophages, dendritic cells, Langerhans's cells, neutrophils, basophils, mast cells, and on some activated T cells and NK cells. CD33 also serves as a key marker and target expressed on acute myeloid leukemia (AML) cells. The long CD33 isoform consists of an N-terminal IgV domain, the sialic acid-binding region, followed by an IgC2 domain, a transmembrane region, and a cytoplasmic domain with one immunoreceptor tyrosine-based inhibitory motifs (ITIM) and one ITIM-like motif in its cytoplasmic domain. The shorter CD33 isoform lacks the ligand-binding IgV domain. The IgV domain plays roles in ligand binding and cellular adhesion whereas the cytoplasmic ITIM tails enable CD33 to transduce inhibitory signaling that can regulate cellular functions such as phagocytosis and cytokine production. Among its two isoforms, the long form contains an IgV domain required for ligand binding, eg, sialylated glycoproteins and gangliosides, whereas the short form lacks this domain. Gemtuzumab297.rMAb binds the IgV domain and therefore detects only the long but not short CD33 isoform as the WM53 and P67.6 antibodies also reportedly do. The HIM3-4 antibody reportedly recognizes both CD33 isoforms. The Gemtuzumab297.rMAb is intended for research use only. It is not intended for use in therapeutic or diagnostic procedures for humans or animals.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
7. An isotype control should be used at the same concentration as the antibody of interest.
8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
11. For U.S. patents that may apply, see bd.com/patents.
