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      RB744 Mouse Anti-Human CD62L (L-Selectin)
      RB744 Mouse Anti-Human CD62L (L-Selectin)

      Multiparameter flow cytometric analysis of CD62L (L-Selectin) expression on Human peripheral blood leukocyte populations.  Human whole blood was first treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and then preincubated with BD Pharmingen™ Human BD Fc Block\"\" (Cat. No. 564219/564220). The leukocytes were then stained with either BD Horizon™ RB744 Mouse IgG1, κ Isotype Control (Cat. No. 570519; Left Plot) or BD Horizon™ RB744 Mouse Anti-Human CD62L (L-Selectin) antibody (Cat. No. 570595/570683; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD62L (L-Selectin) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software.

      RB744 Mouse Anti-Human CD62L (L-Selectin)

      Multiparameter flow cytometric analysis of CD62L (L-Selectin) expression on Human peripheral blood leukocyte populations.  Human whole blood was first treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and then preincubated with BD Pharmingen™ Human BD Fc Block\"\" (Cat. No. 564219/564220). The leukocytes were then stained with either BD Horizon™ RB744 Mouse IgG1, κ Isotype Control (Cat. No. 570519; Left Plot) or BD Horizon™ RB744 Mouse Anti-Human CD62L (L-Selectin) antibody (Cat. No. 570595/570683; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD62L (L-Selectin) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software.

      Product Details
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      BD Horizon™
      SELL; L-selectin; LSEL; LAM-1; LECAM-1; LEU8; LNHR; MEL-14; PLNHR; TQ-1
      Human (QC Testing)
      Mouse IgG1, κ
      Supernatant from PMA-activated Human Peripheral Blood Leukocytes
      Flow cytometry (Routinely Tested)
      5 µl/test
      V S056
      6402
      AB_3685875
      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.
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      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.

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      Product Notices

      1. 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.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      3. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
      4. An isotype control should be used at the same concentration as the antibody of interest.
      5. 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.
      6. 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.
      7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      8. 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.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. For U.S. patents that may apply, see bd.com/patents.
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      570683 Rev. 1
      Antibody Details
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      DREG-56

      The  DREG-56 monoclonal antibody specifically binds to CD62L. CD62L is a 76-95 kDa glycoprotein that is also referred to as L-selectin or LECAM-1. CD62L is expressed on neutrophils, monocytes, T- and B-lymphocyte subsets and NK cells. The DREG-56 antibody recognizes the same antigen as LAM-1, and specifically inhibits >90% of binding of human lymphocytes to high endothelial venules (HEV) in frozen sections of peripheral, but not mucosal lymphoid tissue. It thus defines L-selectin as a human lymphocyte homing receptor for peripheral lymph node HEV.

      570683 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).
      altImg
      RB744
      Blue 488 nm
      498 nm
      746 nm
      570683 Rev.1
      Citations & References
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      View product citations for antibody "570683" on CiteAb

      Development References (5)

      1. Ivetic A, Hoskins Green HL, Hart SJ. L-selectin: A Major Regulator of Leukocyte Adhesion, Migration and Signaling.. Front Immunol. 2019; 10:1068. (Biology). View Reference
      2. Kishimoto TK, Jutila MA, Butcher EC. Identification of a human peripheral lymph node homing receptor: a rapidly down-regulated adhesion molecule. Proc Natl Acad Sci U S A. 1990; 87(6):2244-2248. (Clone-specific: Inhibition). View Reference
      3. Kishimoto TK, Warnock RA, Jutila MA, et al. Antibodies against human neutrophil LECAM-1 (LAM-1/Leu-8/DREG-56 antigen) and endothelial cell ELAM-1 inhibit a common CD18-independent adhesion pathway in vitro. Blood. 1991; 78(3):805-811. (Immunogen: Flow cytometry, Inhibition). View Reference
      4. Polley MJ, Phillips ML, Wayner E, et al. CD62 and endothelial cell-leukocyte adhesion molecule 1 (ELAM-1) recognize the same carbohydrate ligand, sialyl-Lewis x. Proc Natl Acad Sci U S A. 1991; 88(14):6224-6228. (Biology). View Reference
      5. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
      View All (5) View Less
      570683 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.