The 3D3 monoclonal antibody specifically recognizes FcγRII (CD32), a 40 kDa, polymorphic type I transmembrane glycoprotein that serves as a low affinity receptor for aggregated IgG. This highly glycosylated molecule (encoded by at least two different genes) is expressed on monocytes, granulocytes, platelets and B cells. Unlike the FLI28.26 mAb, the 3D3 mAb detected a polymorphic CD32 antigen expressed on B cells of all donors, but only on platelets, monocytes and granulocytes of some donors. The platelets from 3D3+ donors respond to certain stimulatory mAb such as CD165 (clone SN2) which results in aggregation. On the other hand, the platelets from 3D3 negative donors do not form aggregates after stimulation. Individuals can be divided into two groups as responder and non-responder depending on expression, or non-expression, of 3D3. In comparison to the 3D3 mAb, the FLI8.26 mAb detects a monomorphic CD32 antigen expressed on all human donors.
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.
1. Researchers should determine the optimal concentration of this reagent for their individual applications.
2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
3. An isotype control should be used at the same concentration as the antibody of interest.
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. Alexa Fluor® is a registered trademark of Life Technologies Corporation.
6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
8. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email:
[email protected].
