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RB744 Rat Anti-Mouse Ly-49D
Product Details
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BD OptiBuild™
Ly49d; Ly-49d; Klra4; Chok
Mouse (Tested in Development)
Rat F344, also known as Fischer, CDF IgG2a, κ
Not Reported
Flow cytometry (Qualified)
0.2 mg/ml
16635
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


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 to the dye under optimum conditions that minimize unconjugated dye and antibody.

Recommended Assay Procedures

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.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  3. For U.S. patents that may apply, see bd.com/patents.
  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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. 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.
  7. 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.
  8. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. An isotype control should be used at the same concentration as the antibody of interest.
  11. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
757096 Rev. 1
Antibody Details
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4E5

The 4E5 monoclonal antibody specifically recognizes Ly-49D, which is expressed on subsets of natural killer (NK) cells in C57BL/6, C3H/He (at a very low frequency), and SJL, but not DBA/2, AKR, CBA/J, or BALB/c mice. Unlike Ly-49D antigen has not been betected on NK-1.1+ (or DX5+) T cells. In 129/J mice, the 4E5 antibody cross-reacts with Ly-49O, Ly-49R, and Ly-49V. 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. Ly-49D weakly binds to MHC class I antigens of the k halpotype, and Ly-49D+ IL-2-activated NK cells lyse target cells expressing H-2[a], H-2[b], H-2[d], H-2[k], H-2[p], H-2[q], and H-2[s] and the CHO (Chinese hamster ovary) cell line. Ly-49D+ cells mediate allogenic resistance to H-2d bone marrow transplantation. In vitro studies suggest that the Ly-49D receptor mediates activation of NK-cell cytolytic activity via tyrosine phosphorylation of their ITIMs (Immunoreceptor Tyrosine-based Inhibitory Motifs). Molecular differences between the Ly-49D stimulatory receptor and the inhibitory members of the Ly-49 family include the absence of an ITIM in Ly-49D, the lack of phosphorylation of Ly-49D in activated NK cells, and the association of a novel tyrosine-phosphorylated protein (pp16) with Ly-49D in activated NK cells. Ly-49O and Ly-49V are closely related to Ly-49A[B6] and, like Ly-49A, have ITIM domains. Ly-49O- and Ly-49V-transfected 293T (human kidney epithelial) cells bind tetramers of H-2D[b], D[b], D[k], and L[d]. In addition, the Ly-49V-transfected cells also bind K[b], K[d], and K[k]. Ly-49R is closely related to Ly-49D[B6] and is putative activating receptor due to its  lack of an ITIM domain. Ly-49R-transfected 293T cells bind soluble tetramers of H-2D[b], D[d], D[k], and L[d].

757096 Rev. 1
Format Details
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RB744
The BD Horizon RealBlue™ 744 (RB744) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 746-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB744 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), we recommend using an optical filter centered near 750-nm (e.g., a 750/60-nm bandpass filter).
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RB744
Blue 488 nm
498 nm
746 nm
757096 Rev.1
Citations & References
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View product citations for antibody "757096" on CiteAb

Development References (13)

  1. George TC, Mason LH, Ortaldo JR, Kumar V, Bennett M. Positive recognition of MHC class I molecules by the Ly49D receptor of murine NK cells. J Immunol. 1999; 162(4):2035-2043. (Biology). View Reference
  2. Hanke T, Takizawa H, McMahon CW, et al. Direct assessment of MHC class I binding by seven Ly49 inhibitory NK cell receptors. Immunity. 1999; 11(1):67-77. (Biology). View Reference
  3. Idris AH, Smith HR, Mason LH, Ortaldo JR, Scalzo AA, Yokoyama WM. The natural killer gene complex genetic locus Chok encodes Ly-49D, a target recognition receptor that activates natural killing. Proc Natl Acad Sci U S A. 1999; 96(11):6330-6335. (Biology). View Reference
  4. Makrigiannis AP, Pau AT, Saleh A, Winkler-Pickett R, Ortaldo JR, Anderson SK. Class I MHC-binding characteristics of the 129/J Ly49 repertoire. J Immunol. 2001; 166(8):5034-5043. (Biology). View Reference
  5. Mason LH, Anderson SK, Yokoyama WM, Smith HR, Winkler-Pickett R, Ortaldo JR. The Ly-49D receptor activates murine natural killer cells. J Exp Med. 1996; 184(6):2119-2128. (Biology). View Reference
  6. Mason LH, Gosselin P, Anderson SK, Fogler WE, Ortaldo JR, McVicar DW. Differential tyrosine phosphorylation of inhibitory versus activating Ly-49 receptor proteins and their recruitment of SHP-1 phosphatase. J Immunol. 1997; 159(9):4187-4196. (Biology). View Reference
  7. Mason LH, Willette-Brown J, Anderson SK, et al. Characterization of an associated 16-kDa tyrosine phosphoprotein required for Ly-49D signal transduction. J Immunol. 1998; 160(9):4148-4152. (Biology). View Reference
  8. Ortaldo JR, Mason AT, Winkler-Pickett R, Raziuddin A, Murphy WJ, Mason LH. Ly-49 receptor expression and functional analysis in multiple mouse strains. J Leukoc Biol. 1999; 66(3):512-520. (Clone-specific). View Reference
  9. Ortaldo JR, Winkler-Pickett R, Mason AT, Mason LH. The Ly-49 family: regulation of cytotoxicity and cytokine production in murine CD3+ cells. J Immunol. 1998; 160(1):1158-1165. (Clone-specific). View Reference
  10. Raulet DH, Held W, Correa I, Dorfman JR, Wu MF, Corral L. Specificity, tolerance and developmental regulation of natural killer cells defined by expression of class I-specific Ly49 receptors. Immunol Rev. 1997; 155:41-52. (Biology). View Reference
  11. Raziuddin A, Longo DL, Mason L, Ortaldo JR, Bennett M, Murphy WJ. Differential effects of the rejection of bone marrow allografts by the depletion of activating versus inhibiting Ly-49 natural killer cell subsets. J Immunol. 1998; 160(1):87-94. (Biology). View Reference
  12. Takei F, Brennan J, Mager DL. The Ly-49 family: genes, proteins and recognition of class I MHC. Immunol Rev. 1997; 155:67-77. (Biology). View Reference
  13. Yokoyama WM, Seaman WE. The Ly-49 and NKR-P1 gene families encoding lectin-like receptors on natural killer cells: the NK gene complex. Annu Rev Immunol. 1993; 11:613-635. (Biology). View Reference
View All (13) View Less
757096 Rev. 1

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