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APC-Cy™7 Rat Anti-Mouse CD62L
APC-Cy™7 Rat Anti-Mouse CD62L

Analysis of CD62L on mouse bone marrow.  Bone marrow cells from C57BL/6 mice were stained with the APC-Cy™7 Rat Anti-Mouse CD62L antibody (unshaded) or with a APC-Cy7 Rat IgG2a, κ isotype control (shaded).  Histograms were derived from gated events based on light scattering characteristics for bone marrow.  Flow cytometry was performed on a BD™ LSRII flow cytometry system.

Analysis of CD62L on mouse bone marrow.  Bone marrow cells from C57BL/6 mice were stained with the APC-Cy™7 Rat Anti-Mouse CD62L antibody (unshaded) or with a APC-Cy7 Rat IgG2a, κ isotype control (shaded).  Histograms were derived from gated events based on light scattering characteristics for bone marrow.  Flow cytometry was performed on a BD™ LSRII flow cytometry system.

Product Details
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BD Pharmingen™
Sell; L-selectin; LECAM-1; LAM-1; Lnhr; Ly-22; Ly-m22; Lyam-1
Mouse (QC Testing)
Rat F344, also known as Fischer, CDF IgG2a, κ
C3H/eb mouse B lymphoma 38C-13
Flow cytometry (Routinely Tested)
0.2 mg/ml
20343
AB_10611861
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 APC-Cy7 under optimum conditions, and unconjugated antibody and free APC-Cy7 were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  4. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
  5. APC-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher.
  6. APC-Cy7 is a tandem fluorochrome composed of Allophycocyanin (APC), which is excited by laser lines between 595 and 647 nm and serves as an energy donor, coupled to the cyanine dye Cy7™, which acts as an energy acceptor and fluoresces at 780 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in APC-Cy7, thus maximizing its fluorescence emission intensity, minimizing residual emission from APC, and minimizing required electronic compensation in multilaser-laser flow cytometry systems. Note: Although every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-Cy7 conjugate.
  7. Cy is a trademark of GE Healthcare.
  8. 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.
  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.
560514 Rev. 3
Antibody Details
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MEL-14

The MEL-14 monoclonal antibody specifically binds to CD62L (L-selectin), a 95 kDa (on neutrophils) or 74 kDa (on lymphocytes) receptor with lectin-like and Epidermal Growth Factor-like domains. In the mouse, L-selectin is detected on most thymocytes, with the highest levels of expression on an immunocompetent subset and a population of dividing progenitor cells, and on peripheral leukocytes, including subsets of B and T lymphocytes, neutrophils, monocytes, and eosinophils. This member of the selectin adhesion molecule family appears to be required for lymphocyte homing to peripheral lymph nodes and to contribute to neutrophil emigration at inflammatory sites. L-selectin is rapidly shed from lymphocytes and neutrophils upon cellular activation; metalloproteinases may mediate the release of CD62L ectodomains from the cell surface. The level of CD62L expression, along with other markers, distinguishes naive, effector, and memory T cells. L-selectin binds to sialytaed oligosaccharide determinants on high endothelial venules (HEV) in peripheral lymph nodes. In vitro studies have demonstrated that CD34, GlyCAM-1, and MAdCAM-1, all recognized by mAb MECA-79 (anti-mouse PNAd Carbohydrate Epitope, Cat. No. 553863), may be ligands for CD62L. MEL-14 mAb blocks in vitro binding of lymphocytes to peripheral lymph node HEV and inhibits in vivo lymphocyte extravasation into peripheral lymph nodes and late stages of leukocyte rolling.

  

560514 Rev. 3
Format Details
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APC-Cy7
APC-Cy7 dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of a Allophycocyanin (APC) donor that has excitation maxima (Ex Max) of 651 nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 779 nm. APC-Cy7 can be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 780 nm (e.g., a 760/60 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC-Cy7
Red 627-640 nm
651 nm
779 nm
560514 Rev.3
Citations & References
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Development References (20)

  1. Cerwenka A, Carter LL, Reome JB, Swain SL, Dutton RW. In vivo persistence of CD8 polarized T cell subsets producing type 1 or type 2 cytokines. J Immunol. 1998; 161(1):97-105. (Biology). View Reference
  2. Gallatin WM, Weissman IL, Butcher EC. A cell-surface molecule involved in organ-specific homing of lymphocytes. Nature. 1983; 304(5921):30-34. (Immunogen). View Reference
  3. Iwabuchi K, Ohgama J, Ogasawara K, et al. Distribution of MEL-14+ cells in various lymphoid tissues. Immunobiology. 1991; 182(2):161-173. (Biology). View Reference
  4. Jung TM, Gallatin WM, Weissman IL, Dailey MO. Down-regulation of homing receptors after T cell activation. J Immunol. 1988; 141(12):4110-4117. (Biology). View Reference
  5. Kishimoto TK, Jutila MA, Berg EL, Butcher EC. Neutrophil Mac-1 and MEL-14 adhesion proteins inversely regulated by chemotactic factors. Science. 1989; 245(4923):1238-1241. (Biology). View Reference
  6. Lanzavecchia A, Sallusto F. Dynamics of T lymphocyte responses: intermediates, effectors, and memory cells. Science. 2000; 290(5489):92-97. (Biology). View Reference
  7. Lewinsohn DM, Bargatze RF, Butcher EC. Leukocyte-endothelial cell recognition: evidence of a common molecular mechanism shared by neutrophils, lymphocytes, and other leukocytes. J Immunol. 1987; 138(12):4313-4321. (Biology). View Reference
  8. Ley K, Bullard DC, Arbones ML, et al. Sequential contribution of L- and P-selectin to leukocyte rolling in vivo. J Exp Med. 1995; 181(2):669-675. (Biology). View Reference
  9. Mobley JL, Dailey MO. Regulation of adhesion molecule expression by CD8 T cells in vivo. I. Differential regulation of gp90MEL-14 (LECAM-1), Pgp-1, LFA-1, and VLA-4 alpha during the differentiation of cytotoxic T lymphocytes induced by allografts. J Immunol. 1992; 148(8):2348-2356. (Biology). View Reference
  10. Peschon JJ, Slack JL, Reddy P, et al. An essential role for ectodomain shedding in mammalian development. Science. 1998; 282(5392):1281-1284. (Biology). View Reference
  11. Pizcueta P, Luscinskas FW. Monoclonal antibody blockade of L-selectin inhibits mononuclear leukocyte recruitment to inflammatory sites in vivo. Am J Pathol. 1994; 145(2):461-469. (Biology). View Reference
  12. Reichert RA, Jerabek L, Gallatin WM, Butcher EC, Weissman IL. Ontogeny of lymphocyte homing receptor expression in the mouse thymus. J Immunol. 1986; 136(10):3535-3542. (Biology). View Reference
  13. Reichert RA, Weissman IL, Butcher EC. Dual immunofluorescence studies of cortisone-induced thymic involution: evidence for a major cortical component to cortisone-resistant thymocytes. J Immunol. 1986; 136(10):3529-3534. (Biology). View Reference
  14. Reichert RA, Weissman IL, Butcher EC. Phenotypic analysis of thymocytes that express homing receptors for peripheral lymph nodes. J Immunol. 1986; 136(10):3521-3528. (Biology). View Reference
  15. Seibold F, Seibold-Schmid B, Cong Y, et al. Regional differences in L-selectin expression in murine intestinal lymphocytes. Gastroenterology. 1998; 114(5):965-974. (Biology). View Reference
  16. Shortman K, Wilson A, Van Ewijk W, Scollay R. Phenotype and localization of thymocytes expressing the homing receptor-associated antigen MEL-14: arguments for the view that most mature thymocytes are located in the medulla. J Immunol. 1987; 138(2):342-351. (Biology). View Reference
  17. Siegelman MH, Cheng IC, Weissman IL, Wakeland EK. The mouse lymph node homing receptor is identical with the lymphocyte cell surface marker Ly-22: role of the EGF domain in endothelial binding. Cell. 1990; 61(4):611-622. (Biology). View Reference
  18. Sprent J, Tough DF. Lymphocyte life-span and memory. Science. 1994; 265(5177):1395-1400. (Biology). View Reference
  19. Vestweber D. Ligand-specificity of the selectins. J Cell Biochem. 1996; 61(4):585-591. (Biology). View Reference
  20. Yang G, Mizuno MT, Hellstrom KE, Chen L. B7-negative versus B7-positive P815 tumor: differential requirements for priming of an antitumor immune response in lymph nodes. J Immunol. 1997; 158(2):851-858. (Biology). View Reference
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560514 Rev. 3

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