Description
The 4D11 antibody specifically recognizes Ly-49G2 (also known as LGL-1), an inhibitory receptor which is expressed on subsets of natural killer (NK) cells and DX5-positive T lymphocytes (NK-T cells) in all strains tested (e.g., AKR/N, BALB/c, C3H/HeJ, C57BL/6, CBA/J, DBA/2, SJL, 129) and on a population of memory CD8+ T lymphocytes in C57BL/6 mice. Cross-reaction of 4D11 antibody to Ly-49A[B6], Ly-49A[BALB], and Ly-49T[129/J] inhibitory receptors and Ly-49L[CBA/J] activating receptor has been reported. The proportion of NK-T cells expressing Ly-49A and Ly-49G2 is higher (2-5 fold) in thymus than in liver (immature and mature NK-T cells, respectively), and there is evidence that down-regulation of Ly-49 receptor expression is necessary for normal NK-T-cell development to occur. Most NK cells express a single allele of Ly-49A and/or Ly-49G2, although occasionally they may express more than one allele. The Ly-49 family of NK-cell receptors, members of the C-type lectin superfamily, are disulfide-linked type-II transmembrane protein homodimers with extracellular carbohydrate-recognition domains, which bind to MHC class I alloantigens. The Ly-49 family members are expressed independently, such that an individual NK or T cell may display more than one class of Ly-49 receptor homodimers. Binding of Ly-49G[B6]-expressing transfectants to H-2Dd+/H-2Ld+ ConA blasts has been demonstrated, and H-2D[d]-expressing target cells inhibit the lytic activity of Ly-49G2-expressing NK cells. The levels of the Ly-49 inhibitory receptors are down-regulated by their ligands in vivo, and the various levels of expression of a Ly-49 inhibitory receptor may affect the specificity of NK cells. Ly-49G2[+] NK cells are able to lyse target tumor cells expressing H-2[a] and H-2[b] MHC class I antigens in vitro, and they mediate allogeneic and hybrid resistance to H-2[b] bone marrow transplantation. The Ly-49A[BALB] and Ly-49A[B6] alloantigens bind to MHC class I antigens of the d and k haplotypes, and Ly-49A[+] IL-2-activated NK cells are unable to lyse target cells expressing H-2[d] and H-2[k]. In vitro studies suggest that the Ly-49G2 and Ly-49A receptors mediate negative regulation of NK-cell cytolytic activity via tyrosine phosphorylation of their ITIMs (Immunoreceptor Tyrosine-based Inhibitory Motifs). Ly-49T[129/J] has a unique ITIM sequence, and Ly-49T-transfected 293T (human kidney epithelial) cells do not bind soluble tetramers of any tested H-2 alloantigen (D[b], D[d], D[k], K[b], K[d], K[k], L[d]).
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.
Format
- FormatAntibody-Oligo
Suggested Companion Products
Stain Buffer (FBS) RUO 500mLCat No: 554656
Single-Cell Analysis SystemRUO 1EachCat No: 633701
Express Single-Cell Analysis System Package 1EACat No: 633707
Targeted mRNA and AbSeq Training Kit 4 Pack 1EACat No: 633772
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) 2.4G2RUO 0.1mgCat No: 553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) 2.4G2RUO 0.5mgCat No: 553142
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Preparation and Storage
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 and conjugated to BD AbSeq oligonucleotide under optimal conditions.
Product Notices
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- 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.
- Illumina is a trademark of Illumina, Inc.
- This product is covered by one or more of the following patents: US 8,835,358; US 9,290,808; US 9,290,809; US 9,315,857; US 9,567,645; US 9,567,646; US 9,598,736; US 9,708,659; and US 9,816,137.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. Diagnostic uses require a separate license.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to bd.com/genomics-resources for technical protocols.
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.
