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PE Rat Anti-Mouse Ly-49A
PE Rat Anti-Mouse Ly-49A
Flow cytometric analysis of Ly-49A expression on Mouse splenocytes. BALB/c Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with APC Rat Anti-Mouse CD49b antibody (Cat No. 560628) and with either PE Rat IgG2b, κ Isotype Control (Cat. No. 555848; Left Plot) or PE Mouse Anti-Rat Ly-49A antibody (Cat. No. 569156/569157; Right Plot) at 0.06 μg/test. 7-AAD (7-Amino-Actinomycin D) Solution (Cat. No. 559925) was added to cells right before analysis. A bivariate pseudocolor density plot showing the correlated expression of Ly-49A (or Ig Isotype control staining) versus CD49b was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of Ly-49A expression on Mouse splenocytes. BALB/c Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with APC Rat Anti-Mouse CD49b antibody (Cat No. 560628) and with either PE Rat IgG2b, κ Isotype Control (Cat. No. 555848; Left Plot) or PE Mouse Anti-Rat Ly-49A antibody (Cat. No. 569156/569157; Right Plot) at 0.06 μg/test. 7-AAD (7-Amino-Actinomycin D) Solution (Cat. No. 559925) was added to cells right before analysis. A bivariate pseudocolor density plot showing the correlated expression of Ly-49A (or Ig Isotype control staining) versus CD49b was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
Klra1; lymphocyte antigen 49a; Ly-49a; Ly49a; YE1/48; A1
Mouse (QC Testing)
Rat F344, also known as Fischer, CDF IgG2b
ECA17.9.8 T Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
16627
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 and unconjugated antibody and free dye were removed.

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 cell 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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
569157 Rev. 1
Antibody Details
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YE1/48.10.6

The YE1/48.10.6 monoclonal antibody specifically recognizes Lymphocyte antigen 49a (Ly-49A). Ly-49A is a single-pass type-II transmembrane that is encoded by Klra1 (Killer cell lectin-like receptor, subfamily A member 1) which belongs to the C-type lectin superfamily. Ly-49A is comprised of an extracellular carbohydrate-recognition domain (CRD) that can bind strongly to H-2D MHC class I alloantigens expressed on target cells including H-2D[d], H-2D[k], and H-2D[p]. The extracellular domain is followed by a transmembrane region and a cytoplasmic domain with an immunoreceptor tyrosine-based inhibitory motif (ITIM). Ly-49A is expressed as a  disulfide-linked homodimer on a subset of NK and NK-T cells. MHC Class I ligand-bound Ly-49A can negatively regulate NK-cell cytolytic activity via tyrosine phosphorylation of its ITIM.

569157 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
569157 Rev.1
Citations & References
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Development References (6)

  1. Chan PY, Takei F. Expression of a T cell receptor-like molecule on normal and malignant murine T cells detected by rat monoclonal antibodies to nonclonotypic determinants.. J Immunol. 1986; 136(4):1346-53. (Immunogen: Blocking, Flow cytometry, Immunoprecipitation, Western blot). View Reference
  2. 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: Blocking). View Reference
  3. Nagasawa R, Gross J, Kanagawa O, et al. Identification of a novel T cell surface disulfide-bonded dimer distinct from the alpha/beta antigen receptor. J Immunol. 1987; 138(3):815-824. (Biology). View Reference
  4. Nakamura MC, Seaman WE. Ligand interactions by activating and inhibitory Ly-49 receptors.. Immunol Rev. 2001; 181:138-48. (Biology). View Reference
  5. Takei F. Two surface antigens expressed on proliferating mouse T lymphocytes defined by rat monoclonal antibodies.. J Immunol. 1983; 130(6):2794-7. (Immunogen: Flow cytometry, Radioimmunoassay, Western blot). View Reference
  6. Zimmer J, Ioannidis V, Held W. H-2D ligand expression by Ly49A+ natural killer (NK) cells precludes ligand uptake from environmental cells: implications for NK cell function.. J Exp Med. 2001; 194(10):1531-9. (Clone-specific: Flow cytometry). View Reference
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569157 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.