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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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Since applications vary, each investigator should titrate the reagent to obtain optimal results.
An isotype control should be used at the same concentration as the antibody of interest.
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.
Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
The HRL1 monoclonal antibody specifically binds to L-selectin (LECAM-1, CD62L), which is detected on a small percentage of thymocytes and on most neutrophils and peripheral lymphocytes. CD62L is a 62-kDa (on neutrophils) or 65-kDa (on lymphocytes) receptor, with lectin-like and epidermal growth factor-like domains, which binds to sialylated oligosaccharide determinants on high endothelial venules (HEV) in peripheral lymph nodes. 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 cell activation. In the mouse, the level of CD62L expression distinguishes naive CD4+T cells from effector/memory T helper cells. HRL1 antibody inhibits the ligand-binding activity of Lselectin in in vitro assays and slows in vivo leukocyte rolling on microvascular endothelium.
551398 Rev. 13
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
Citations & References
Mulligan MS, Miyasaka M, Tamatani T, Jones ML, Ward PA. Requirements for L-selectin in neutrophil-mediated lung injury in rats. J Immunol. 1994; 152(2):832-840. (Biology).
Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994; 76(2):301-314. (Biology).
Tamatani T, Kitamura F, Kuida K, et al. Characterization of rat LECAM-1 (L-selectin) by the use of monoclonal antibodies and evidence for the presence of soluble LECAM-1 in rat sera. Eur J Immunol. 1993; 23(9):2181-2188. (Immunogen: Flow cytometry).