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      BB700 Mouse Anti-Human CD58 (LFA-3)
      Product Details
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      BD OptiBuild™
      LFA-3; LFA3; Lymphocyte function-associated antigen 3; Ag3
      Human (Tested in Development)
      Mouse IgG2a, κ
      Lc-LFA-3 mouse L-cell transfectant
      Flow cytometry (Qualified)
      0.2 mg/ml
      V πNK15
      965
      AB_2743301
      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 with BD Horizon BB700 under optimal conditions that minimize unconjugated dye and antibody.
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      Recommended Assay Procedures

      For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes 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 or 566349).

      When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

      For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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      Product Notices

      1. This antibody was developed for use in flow cytometry.
      2. 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.
      3. Researchers should determine the optimal concentration of this reagent for their individual applications.
      4. An isotype control should be used at the same concentration as the antibody of interest.
      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 www.bdbiosciences.com/us/s/resources for technical protocols.
      8. 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.
      9. BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
      10. Cy is a trademark of GE Healthcare.
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      745860 Rev. 1
      Antibody Details
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      L306.4

      The L306.4  monoclonal antibody specifically recognizes CD58 which is also known as Lymphocyte function-associated antigen-3 (LFA-3) or Ag3. CD58 is a ~40 to 65 kDa cell-surface glycoprotein that belongs to the immunoglobulin superfamily. The CD58 antigen mediates cellular adhesion and participates in signal transduction when it binds to its ligand, the CD2 antigen. Cellular interactions regulated by the CD58/CD2 antigens are involved in the antigen-independent adhesion pathway and cytotoxic T lymphocyte (CTL) activity. The CD58 antigen has two isoforms. One isoform is anchored in the cell membrane by a glycophosphatidyl inositol tail, while the other is a type I transmembrane glycoprotein which has a transmembrane hydrophobic segment and a cytoplasmic segment composed of 12 amino acids. The CD58 antigen is widely distributed on cells of both hematopoietic and nonhematopoietic origin. The CD58 antigen is expressed on approximately 40% to 60% of peripheral blood lymphocytes, including CTL. It is also expressed on monocytes, granulocytes, B lymphoblastoid cell lines (such as JY and Daudi), platelets, vascular endothelium and smooth muscle, fibroblasts, and approximately 40% of bone marrow cells.

      The antibody was conjugated to BD Horizon™ BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes.   It is a polymer-based tandem dye developed exclusively by BD Biosciences.  With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

      745860 Rev. 1
      Format Details
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      BB700
      The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BB700
      Blue 488 nm
      476 nm
      695 nm
      745860 Rev.1
      Citations & References
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      Development References (12)

      1. Rincón J, Patarroyo M. Effect of antibodies from the T-cell (‘CD2 only’) and the NK/non-lineage (new panel only) sections on adhesion of Jurkat (T) cells to human erythrocytes. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:718-720.
      2. Carpén O, Dustin ML, Springer TA, Swafford JA, Beckett LA, Caulfield JP. Motility and ultrastructure of large granular lymphocytes on lipid bilayers reconstituted with adhesion receptors LFA-1, ICAM-1, and two isoforms of LFA-3.. J Cell Biol. 1991; 115(3):861-71. (Biology). View Reference
      3. Dengler TJ, Hoffmann JC, Knolle P, et al. Structural and functional epitopes of the human adhesion receptor CD58 (LFA-3). Eur J Immunol. 1992; 22(11):2809-2817. (Biology). View Reference
      4. Griffin H, Rowe M, Murray R, Brooks J, Rickinson A. Restoration of the LFA-3 adhesion pathway in Burkitt's lymphoma cells using an LFA-3 recombinant vaccinia virus: consequences for T cell recognition.. Eur J Immunol. 1992; 22(7):1741-8. (Biology). View Reference
      5. Krensky AM, Robbins E, Springer TA, Burakoff SJ. LFA-1, LFA-2, and LFA-3 antigens are involved in CTL-target conjugation.. J Immunol. 1984; 132(5):2180-2. (Biology). View Reference
      6. Krensky AM, Sanchez-Madrid F, Robbins E, Nagy JA, Springer TA, Burakoff SJ. The functional significance, distribution, and structure of LFA-1, LFA-2, and LFA-3: cell surface antigens associated with CTL-target interactions.. J Immunol. 1983; 131(2):611-6. (Biology). View Reference
      7. Scheeren RA, Koopman G, Van der Baan S, Meijer CJ, Pals ST. Adhesion receptors involved in clustering of blood dendritic cells and T lymphocytes.. Eur J Immunol. 1991; 21(5):1101-5. (Biology). View Reference
      8. Selvaraj P, Plunkett ML, Dustin M, Sanders ME, Shaw S, Springer TA. The T lymphocyte glycoprotein CD2 binds the cell surface ligand LFA-3.. Nature. 326(6111):400-3. (Biology). View Reference
      9. Shaw S, Johnson JP. Cluster report: CD58. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:714-716.
      10. Shaw S, Luce GE, Quinones R, Gress RE, Springer TA, Sanders ME. Two antigen-independent adhesion pathways used by human cytotoxic T-cell clones.. Nature. 323(6085):262-4. (Biology). View Reference
      11. Smith ME, Thomas JA. Cellular expression of lymphocyte function associated antigens and the intercellular adhesion molecule-1 in normal tissue.. J Clin Pathol. 1990; 43(11):893-900. (Clone-specific). View Reference
      12. Springer TA. Adhesion receptors of the immune system. Nature. 1990; 346(6283):425-434. (Biology). View Reference
      View All (12) View Less
      745860 Rev. 1

      Please refer to Support Documents for Quality Certificates


      Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


      Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

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

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