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 contents in the cap/tube threads prior to the first opening.
Please refer to the following CITE-Seq protocols for additional staining instructions:
• Single Cell Labelling with BD® AbSeq Ab-Oligos (1 to 40-plex) (Doc ID: 23-24262)
• Single-Cell Labeling with BD® AbSeq Ab-Oligos (41 to 100 plex) (Doc ID: 23-22314)
• Single Cell Labelling with BD® Single-Cell Multiplexing Kits and BD® AbSeq Ab-Oligos (1 to 40 plex) (Doc ID:23-21339)
• Single-Cell Labeling with BD® Single-Cell Multiplexing Kits and BD® AbSeq Ab-Oligos (41 to 100 plex) (Doc ID: 23-22354)
• Single-Cell Labeling with BD® Flex Single-Cell Multiplexing Kits and BD® AbSeq Ab-Oligos (1 plex to 100 plex) (Doc ID:23-24312)
Sequencing reference files can be made using the BD® AbSeq Panel Generator (https://abseq-ref-gen.genomics.bd.com/).
Use standard laboratory safety protocols. Read and understand the safety data sheets (SDSs) before handling chemicals. To obtain SDSs, go to regdocs.bd.com or contact BD Biosciences technical support at scomix@bd.com.
Warning: All biological specimens and materials contacting them are considered biohazardous. Handle as if capable of transmitting infection and dispose of with proper precautions in accordance with federal, state, and local regulations. Never pipette by mouth. Wear suitable protective clothing, eyewear, and gloves.
Product Notices
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
- This reagent has been pre-diluted for use at the recommended volume per test. Typical use is 2 μl for 1 × 10^6 cells in a 200-μl staining reaction. For best results, close caps securely after use and before storage.
- 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. Follow state and local guidelines when disposing of hazardous waste.
- Please refer to https://www.bdbiosciences.com/en-us/resources/protocols/single-cell-multiomics for technical protocols.
The 5G6 monoclonal antibody specifically recognizes Sialic acid-binding Ig-like lectin 10 (Siglec-10). Siglec-10 is a type I transmembrane glycoprotein encoded by SIGLEC10 that belongs to the SIGLEC (sialic acid binding Ig-like lectin) family within the immunoglobulin superfamily. The extracellular region of Siglec-10 contains five Ig-like domains including an N-terminal V-set domain that contains the sialic acid-binding site and four C2-set domains. Siglec-10 preferentially binds to glycoconjugates containing α2,3- or α2,6-linked sialic acids present on human cell surfaces and may function as an adhesion receptor. Its cytoplasmic region contains two immunoreceptor tyrosine-based inhibitory motifs (ITIM) that upon phosphorylation can bind the SH2 domains of several SH2-containing phosphatases. This CD33-related Siglec has a mouse ortholog designated Siglec-G. Siglec-10 is expressed on B cells, eosinophils, monocytes, and a subset of CD16+CD56- natural killer (NK) cells. It may play a role as an inhibitory receptor for leucocytes involved in innate and adaptive immunity. The 5G6 antibody reportedly did not crossreact with Siglecs 3, 5, 7, 8 and 9.
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 various sequencing platforms.
NOTE: The BD Rhapsody™ Single-Cell Analysis System must be used with the BD Rhapsody™ Express Instrument or BD Rhapsody™ HT Xpress Instrument.
Development References (3)
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Macauley MS, Crocker PR, Paulson JC. Siglec-mediated regulation of immune cell function in disease.. Nat Rev Immunol. 2014; 14(10):653-66. (Biology). View Reference
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Munday J, Kerr S, Ni J, et al. Identification, characterization and leucocyte expression of Siglec-10, a novel human sialic acid-binding receptor.. Biochem J. 2001; 355(Pt 2):489-97. (Immunogen: ELISA, Flow cytometry). View Reference
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Pillai S, Netravali IA, Cariappa A, Mattoo H. Siglecs and immune regulation.. Annu Rev Immunol. 2012; 30:357-92. (Biology). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.