BD™ AbSeq Oligo Hamster Anti-Mouse FcγRIV (CD16-2)
Clone 9E9
Preparation And Storage
Recommended Assay Procedures
Put all BD AbSeq Reagents to be pooled into a Latch Rack for 500 µL Tubes (Thermo Fisher Scientific Cat. No. 4900). Arrange the tubes so that they can be easily uncapped and re-capped with an 8-Channel Screw Cap Tube Capper (Thermo Fisher Scientific Cat. No. 4105MAT) and the reagents aliquoted with a multi-channel pipette.
BD AbSeq tubes should be centrifuged for ≥ 30 seconds at 400 × g to ensure removal of any content in the cap/tube threads prior to the first opening.
The 9E9 monoclonal antibody specifically binds to Fc receptor, IgG, low affinity IV (Fc γ RIV), which is also known as CD16-2. Fc γ RIV is encoded by Fcgr4 and belongs to the family of receptors for the Fc region of IgG (Fc γ Rs), which also includes Fc γ RI (CD64), Fc γ RII (CD32), and Fc γ RIII (CD16) within the Ig superfamily. FcγRIV is expressed on monocytes, macrophages, dendritic cells, and neutrophils. FcγRIV, w hich serves as a ligand-biding subunit, requires the common Fcγ chain for expression and signaling . This complex serves as a cell activating receptor when bound by IgG2a or IgG2b. Fc γ RIV plays various roles in inflammation including neutrophil trafficking and mast cell degranulation. FcγRIV can also function as a low-affinity IgE receptor and promote IgE-induced inflammation. The 9E9 antibody can reportedly inhibit cellular FcγRIV (CD16-2) function and block non-antigen-specific binding of immune com plexes to improve the quality of immunological assays.
Application Notes
The antibody was conjugated to an oligonucleotide that contains an antibody clone-specific barcode (ABC) flanked by a poly-A tail on the 3' end and a PCR handle (PCR primer binding site) on the 5' end. The ABC for this antibody was designed to be used with other BD AbSeq oligonucleotides conjugated to other antibodies. All AbSeq ABC sequences were selected in silico to be unique from human and mouse genomes, have low predicted secondary structure, and have high Hamming distance within the BD AbSeq portfolio, to allow for sequencing error correction and unique mapping. The poly-A tail of the oligonucleotide allows the ABC to be captured by the BD Rhapsody™ system. The 5' PCR handle allows for efficient sequencing library generation for Illumina sequencing platforms.
NOTE: The BD Rhapsody Single-Cell Analysis System must be used with the BD Rhapsody Express Instrument.
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