Fcγ receptor type II (FcγRII) molecules, also known as CD32 antigens, serve as low affinity receptors for monomeric IgG but bind immune complexes (aggregated IgG) efficiently. Several forms of these receptors, including transmembrane or soluble glycoproteins, are encoded by separate genes and by alternative mRNA splicing: CD32a/FcγRIIa (FCGR2A), CD32b/FcγRIIb (FCGR2B), and CD32c/FcγRIIc (FCGR2C). CD32 molecules are comprised of two IgC-like domains that may be followed by a transmembrane region and a cytoplasmic domain with either ITAM (CD32a and CD32c) or ITIM (CD32b) immunoreceptor signaling motifs. These polymorphic receptors are differentially expressed by leucocyte subsets and are involved in the process of phagocytosis, clearing of immune complexes, platelet activation and degranulation, and regulation of immune responses. The IV.3 monoclonal antibody strongly recognizes FcγRIIA expressed on platelets, monocytes, macrophages, neutrophils, eosinophils, basophils, and B cells. It reportedly binds to an epitope mapped to amino acids 132-137 (FSHLDP) located in the second IgC-like domain within the ligand-binding site. The IV.3 antibody has been used to crosslink or block FcγRII in functional studies as well as a blocking antibody to reduce non-specific binding by antibodies used for staining or cell separation applications. This antibody may weakly crossreact with FcγRIIb/CD32b.
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® CompBeads 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 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].
9. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
