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RB744 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
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.
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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.

757042 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
757042 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
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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.