BUV563 Rat Anti-Mouse Ly-49G2
Clone 4D11 (RUO)
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- Alternative Name LGL-1; Klra7
- Concentration 0.2 mg/ml
- Isotype Rat F344, also known as Fischer, CDF IgG2a, κ
- Reactivity Mouse (Tested in Development)
Flow cytometry (Qualified)
- Immunogen Large granular lymphocytes (LGL) enriched from C57BL/6N mouse liver
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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]).
The antibody was conjugated to BD Horizon™ BUV563 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 which has an Ex Max of 348 nm and an acceptor dye. The tandem has an Em Max at 563 nm. BD Horizon BUV563 can be excited by the 355 nm ultraviolet laser. On instruments with a 561 nm Yellow-Green laser, the recommended bandpass filter is 585/15 nm with a 535 nm long pass to minimize laser light leakage. When BD Horizon BUV563 is used with an instrument that does not have a 561 nm laser, a 560/40 nm filter with a 535 nm long pass may be more optimal. Due to the excitation and emission characteristics of the acceptor dye, there may be spillover into the PE and PE-CF594 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
BUV563 is a tandem fluorochrome that combines BD Horizon BUV395 and an acceptor dye with an EM Max at 563 nm. Due to the excitation of the acceptor dye by other laser lines, there may be spillover into channels detecting PE (eg, 575/26-nm filter) and PE-CF594 (eg, 610/20-nm filter). BUV563 has been exclusively developed by BD Biosciences for instruments equipped with a 355-nm UV laser.
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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.
The antibody was conjugated with BD Horizon BUV563 under optimal conditions that minimize unconjugated dye and antibody.
- This antibody was developed for use in flow cytometry.
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).