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Alexa Fluor™ 488 Rat Anti-Mouse Ly-49A

BD Pharmingen™ Alexa Fluor™ 488 Rat Anti-Mouse Ly-49A

Clone YE1/48.10.6 (also known as YE1/48)

(RUO)
Alexa Fluor™ 488 Rat Anti-Mouse Ly-49A
Multicolor flow cytometric analysis of Ly-49A expression on Mouse splenic leukocytes.  C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with BD Horizon™ BV421 Rat Anti-Mouse CD49b antibody (Cat. No. 563063) and with either Alexa Fluor™ 488 Rat IgG2b, κ Isotype Control (Cat. No. 557726; Left Plot) or Alexa Fluor™ 488 Rat Anti-Mouse Ly-49A antibody (Cat. No. 570081/570162; Right Plot) at 0.25 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The 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) splenic leukocytes. 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.
Multicolor flow cytometric analysis of Ly-49A expression on Mouse splenic leukocytes.  C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with BD Horizon™ BV421 Rat Anti-Mouse CD49b antibody (Cat. No. 563063) and with either Alexa Fluor™ 488 Rat IgG2b, κ Isotype Control (Cat. No. 557726; Left Plot) or Alexa Fluor™ 488 Rat Anti-Mouse Ly-49A antibody (Cat. No. 570081/570162; Right Plot) at 0.25 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The 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) splenic leukocytes. 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

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 unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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. 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. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  9. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  10. For U.S. patents that may apply, see bd.com/patents.
570162 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.

570162 Rev. 1
Format Details
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
570162 Rev.1
Citations & References
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View product citations for antibody "570162" on CiteAb

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
View All (6) View Less
570162 Rev. 1

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