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Alexa Fluor® 488 Rat Anti-Mouse CD8a 53-6.7 RUO

Alexa Fluor® 488 Rat Anti-Mouse CD8a 

Clone  53-6.7   (RUO)

  • Brand BD Pharmingen™
  • Alternative Name Cd8a; CD8 alpha chain; Ly-2; Lyt2; Lyt-2; Ly-35; Ly-B
  • Concentration 0.2 mg/ml
  • Isotype Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ
  • Reactivity Mouse (QC Testing) 
  • Application Flow cytometry (Routinely Tested) 
    Immunofluorescence (Tested During Development) 
  • Immunogen Mouse Spleen Cells or Thymocyte Membranes
  • Entrez Gene ID 12525 
  • Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
  • Regulatory Status RUO
Product Details Recommended Assay References

Description

The 53-6.7 monoclonal antibody specifically binds to the 38 kDa α and 34 kDa α' chains of the CD8 differentiation antigen (Ly-2 or Lyt-2) of all mouse strains tested. The CD8 α and α' chains (CD8a) form heterodimers with the CD8 β chain (CD8b, Ly-3, or Lyt-3) on the surface of most thymocytes. A subpopulation of mature T lymphocytes (i.e., MHC class I-restricted T cells, including most T suppressor/cytotoxic cells) expresses almost exclusively the CD8 αβ heterodimer. Subsets of γδ TCR-bearing T cells, intestinal intrapithelial lymphocytes, and dendritic cells express CD8a without CD8b. It has been suggested that the expression of the CD8a/CD8b heterodimer is restricted to T lymphocytes which matured in the thymus or in an extrathymic environment that had been influenced by thymus-initiated neuroendocrine signals. CD8 is an antigen coreceptor on the T-cell surface which interacts with MHC class I molecules on antigen-presenting cells or epithelial cells. It participates in T-cell activation through its association with the T-cell receptor complex and protein tyrosine kinase lck (p56 [lck]). The CD8 α and α' chains arise from alternatively spliced messengers of a single CD8a gene. The longer α form associates with p56 [lck] via a CXCP motif in its cytoplasmic domain, which it shares with CD4, but not with CD8b. The truncated α' chain is unable to associate with p56 [lck], and it may function to attenuate the CD8-mediated costimulatory signal during intrathymic T-cell maturation. In vivo and in vitro treatment with 53-6.7 mAb has reportedly been effective at depleting CD8+ peripheral T lymphocytes. The 53-6.7 antibody has also been reported to cross-react with CD8 α- and α'-like polypeptides on subsets of thymic and peripheral lymphocytes in the Egyptian toad, Bufo regularis.

Format
  • Format Alexa Fluor® 488
  • Excitation Source Blue 488 nm
  • Excitation Max 495 nm
  • Emission Max 519 nm

Alexa Fluor® conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of FITC. However, Alexa Fluor® 488 tends to be brighter and more optimal for intracellular applications. Due to nearly identical excitation and emission properties but different spillover characteristics, FITC and Alexa Fluor® 488 cannot be used simultaneously. Alexa Fluor® 488 exhibits extraordinary photostability, which makes it highly suitable for fluorescence microscopy.

Other Formats →

Suggested Companion Products

Fixation Buffer RUO
100 mL Cat No: 554655

Stain Buffer (FBS) RUO
500 mL Cat No: 554656

Alexa Fluor® 488 Rat IgG2a, κ Isotype Control RUO
0.1 mg Cat No: 557676

DAPI Solution RUO
1 mg Cat No: 564907

Alexa Fluor® 647 Rat Anti-Mouse CD19 1D3 RUO
0.1 mg Cat No: 557684

Stain Buffer (BSA) RUO
500 mL Cat No: 554657

Resources & Tools
 Spectrum Viewer  Panel Designer  Spectrum Viewer  Download TDS  Regulatory Document Website
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 Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  5. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.


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  7. Ledbetter JA, Rouse RV, Micklem HS, Herzenberg LA. T cell subsets defined by expression of Lyt-1,2,3 and Thy-1 antigens. Two-parameter immunofluorescence and cytotoxicity analysis with monoclonal antibodies modifies current views. J Exp Med. 1980; 152(2):280-295. (Biology: Flow cytometry, Immunofluorescence). View reference

  8. Ledbetter JA, Seaman WE, Tsu TT, Herzenberg LA. Lyt-2 and lyt-3 antigens are on two different polypeptide subunits linked by disulfide bonds. Relationship of subunits to T cell cytolytic activity. J Exp Med. 1981; 153(6):1503-1516. (Biology: Flow cytometry, Immunofluorescence). View reference

  9. LeFrancois L. Extrathymic differentiation of intraepithelial lymphocytes: generation of a separate and unequal T-cell repertoire. Immunol Today. 1991; 12(12):436-438. (Biology: Flow cytometry, Immunofluorescence). View reference

  10. Leishman AJ, Naidenko OV, Attinger A, et al. T cell responses modulated through interaction between CD8alphaalpha and the nonclassical MHC class I molecule, TL. Science. 2001; 294(5548):1848-1849. (Biology: Flow cytometry, Immunofluorescence). View reference

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  12. Mitnacht R, Bischof A, Torres-Nagel N, Hunig T. Opposite CD4/CD8 lineage decisions of CD4+8+ mouse and rat thymocytes to equivalent triggering signals: correlation with thymic expression of a truncated CD8 alpha chain in mice but not rats. J Immunol. 1998; 160(2):700-707. (Biology: Flow cytometry, Immunofluorescence). View reference

  13. Murosaki S, Yoshikai Y, Ishida A, et al. Failure of T cell receptor V beta negative selection in murine intestinal intra-epithelial lymphocytes. Int Immunol. 1991; 3(10):1005-1013. (Biology: Flow cytometry, Immunofluorescence). View reference

  14. Nakayama K, Nakayama K, Negishi I, et al. Requirement for CD8 beta chain in positive selection of CD8-lineage T cells. Science. 1994; 263(5150):1131-1133. (Biology: Flow cytometry, Immunofluorescence). View reference

  15. Negm HI, Mansour MH, Saad AH, Abdel Halim RS. Structural characterization of an Lyt-2/3 homolog expressed on Bufo regularis lymphocytes. Comp Biochem Physiol B Biochem Mol Biol. 1996; 113(1):79-87. (Biology: Flow cytometry, Immunofluorescence). View reference

  16. O'Rourke AM, Mescher MF. The roles of CD8 in cytotoxic T lymphocyte function. Immunol Today. 1993; 14(4):183-188. (Biology: Flow cytometry, Immunofluorescence). View reference

  17. Rocha B, Vassalli P, Guy-Grand D. The extrathymic T-cell development pathway. Immunol Today. 1992; 14(3):140-141. (Biology: Flow cytometry, Immunofluorescence). View reference

  18. Sydora BC, Brossay L, Hagenbaugh A, Kronenberg M, Cheroutre H. TAP-independent selection of CD8+ intestinal intraepithelial lymphocytes. J Immunol. 1996; 156(11):4209-4216. (Biology: Flow cytometry, Immunofluorescence). View reference

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  20. van Ewijk W, van Soest PL, van den Engh GJ. Fluorescence analysis and anatomic distribution of mouse T lymphocyte subsets defined by monoclonal antibodies to the antigens Thy-1, Lyt-1, Lyt-2, and T-200. J Immunol. 1981; 127(6):2594-2604. (Biology: Flow cytometry, Immunofluorescence). View reference

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  22. Wang J, Klein JR. Thymus-neuroendocrine interactions in extrathymic T cell development. Science. 1994; 265(5180):1860-1862. (Biology: Flow cytometry, Immunofluorescence). View reference

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