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Multiparameter flow cytometric analysis of IREM-2 (CD300e) expression on human peripheral blood leucocytes. Whole blood cells were stained with either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565357; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human IREM-2 (CD300e) antibody (Cat. No. 566315; Right Plot). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-parameter flow cytometric contour plots showing the correlated expression of IREM-2 (CD300e) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
BD Pharmingen™ Alexa Fluor® 647 Mouse Anti-Human IREM-2 (CD300e)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
Companion Products
The UP-H1 monoclonal antibody recognizes Immune receptor expressed by myeloid cells 2 (IREM-2) which is also known as CD300 antigen like family member E (CD300e), CMRF35-like molecule 2 (CLM2), or Polymeric immunoglobulin receptor 2 (PIgR2). IREM-2 is a 34 kDa type I transmembrane glycoprotein which belongs to the CMFRF35 subfamily of the immunoglobulin (Ig) superfamily. IREM-2 is expressed on monocytes, macrophages and myeloid dendritic cells (mDCs), but not on lymphoid dendritic cells or granulocytes. IREM-2 associates with DAP-12, an ITAM-containing adapter molecule, which provides activating signals. Crosslinking of IREM-2 leads to intracellular calcium mobilization, release of proinflammatory cytokines such as TNF, and increased expression of activation markers on monocytes and mDCs. IREM-2 activation of mDCs can reportedly enhance the alloreactive response of naive T cells.
Development References (3)
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Aguilar H, Alvarez-Errico D, García-Montero AC, Orfao A, Sayós J, López-Botet M. Molecular characterization of a novel immune receptor restricted to the monocytic lineage. J Immunol. 2004; 173(11):6703-6711. (Immunogen: Flow cytometry, Immunoprecipitation, Radioimmunoassay). View Reference
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Borrego F. The CD300 molecules: an emerging family of regulators of the immune system. Blood. 2013; 121(11):1951-1960. (Biology). View Reference
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Brckalo T, Calzetti F, Pérez-Cabezas B, Borràs FE, Cassatella MA, López-Botet M. Functional analysis of the CD300e receptor in human monocytes and myeloid dendritic cells.. Eur J Immunol. 2010; 40(3):722-32. (Clone-specific: Flow cytometry). View Reference
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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.