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PE Mouse Anti-Human CD1e
PE Mouse Anti-Human CD1e
Flow cytometric analysis of CD1e expression in human Jurkat cells. Cells from the human Jurkat (Acute T cell leukemia, ATCC TIB-152) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Perm/Wash Buffer (Cat. No. 554723). Cells were stained with either PE Mouse IgG1, κ Isotype Control (Cat No. 554680, dashed line histogram) or PE Mouse Anti-Human CD1e antibody (Cat No. 567271/567272; solid line histogram) at 0.125 µg/test. The fluorescence histogram showing intracellular CD1e expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact Jurkat cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System equipped with a Yellow-Green Laser and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of CD1e expression in human Jurkat cells. Cells from the human Jurkat (Acute T cell leukemia, ATCC TIB-152) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Perm/Wash Buffer (Cat. No. 554723). Cells were stained with either PE Mouse IgG1, κ Isotype Control (Cat No. 554680, dashed line histogram) or PE Mouse Anti-Human CD1e antibody (Cat No. 567271/567272; solid line histogram) at 0.125 µg/test. The fluorescence histogram showing intracellular CD1e expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact Jurkat cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System equipped with a Yellow-Green Laser and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
CD1E; hCD1e; R2
Human (QC Testing)
Mouse IgG1, κ
Human CD1e Transfected Cell Line
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
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 and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Antibody Details
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20.6

The 20.6 monoclonal antibody specifically recognizes CD1e. Human CD1e is an ~40 kDa single-pass type I transmembrane protein that is encoded by CD1E (CD1e molecule). CD1e belongs to the CD1 family of lipid antigen-presenting, major histocompatibility complex (MHC)-like proteins. CD1e is not expressed on the plasma membrane like the other CD1 molecules and thus does not perform directly as an antigen-presenting molecule. CD1e associates with β2 microglobulin in endosomes and the Golgi before reaching late endosomes and lysosomes. Intracellular CD1e facilitates the processing of self or foreign (eg, microbial) lipids for their presentation to T cells by other CD1 family molecules which are expressed at the cell surface. Human CD1e is constitutively expressed in a limited number of cell types including dendritic cells, Langerhans cells, and thymocytes. Mice and rats lack human CD1e orthologs. Alternate splicing of human CD1e generates multiple isoforms including a soluble form.

        

Format Details
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PE
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.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
Citations & References
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Development References (5)

  1. Angenieux C, Salamero J, Fricker D, et al. Characterization of CD1e, a third type of CD1 molecule expressed in dendritic cells.. J Biol Chem. 2000; 275(48):37757-64. (Immunogen: Flow cytometry, Functional assay, Immunofluorescence). View Reference
  2. Angénieux C, Fraisier V, Maître B, et al. The cellular pathway of CD1e in immature and maturing dendritic cells.. Traffic. 2005; 6(4):286-302. (Clone-specific: Flow cytometry, Functional assay, Immunofluorescence). View Reference
  3. Facciotti F, Cavallari M, Angénieux C, et al. Fine tuning by human CD1e of lipid-specific immune responses.. Proc Natl Acad Sci U S A. 2011; 108(34):14228-33. (Clone-specific: Flow cytometry). View Reference
  4. Tourne S, Maitre B, Collmann A, et al. Cutting edge: a naturally occurring mutation in CD1e impairs lipid antigen presentation.. J Immunol. 2008; 180(6):3642-6. (Clone-specific: Immunofluorescence, Immunoprecipitation). View Reference
  5. de la Salle H, Mariotti S, Angenieux C, et al. Assistance of microbial glycolipid antigen processing by CD1e.. Science. 2005; 310(5752):1321-4. (Clone-specific: Immunoprecipitation). View Reference
View All (5) View Less

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.