Multiparameter flow cytometric analysis of CLM-8 (CD300a) expression on human peripheral blood leucocyte populations. Human whole blood was stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human CLM-8 (CD300a) antibody (Cat. No. 566342; Right Plot) at 0.5 µg/test. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-parameter flow cytometric contour plots showing the correlated expression of CLM-8 (CD300a) [or Ig Isotype control staining] versus side light-scatter signals were derived from gated events with the forward and side light-scattering characteristics of viable leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of CLM-8 (CD300a) expression on human peripheral blood leucocyte populations. Human whole blood was stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human CLM-8 (CD300a) antibody (Cat. No. 566342; Right Plot) at 0.5 µg/test. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-parameter flow cytometric contour plots showing the correlated expression of CLM-8 (CD300a) [or Ig Isotype control staining] versus side light-scatter signals were derived from gated events with the forward and side light-scattering characteristics of viable leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
CD300A; CLM-8; CLM8; CMRF-35-H9; CMRF35H; IGSF12; IRp60; IRC1: IRC2
Human (QC Testing)
Mouse IgG1
HPB Human Acute Lymphoid Leukemia Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2739699
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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Product Notices
- 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.
- 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® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557714
566342 Rev. 2
Antibody Details
MEM-260
The MEM-260 monoclonal antibody specifically recognizes CD300a which is also known as CMRF35-like molecule 8 (CLM8), Inhibitory receptor protein 60 (IRp60), or Immunoglobulin superfamily member 12 (IGSF12). CD300a is a ~60 kDa type I transmembrane glycoprotein that belongs to the CMRF family within the Ig gene superfamily. CD300a is expressed on natural killer (NK) cells, T and B cell subsets, monocytes, macrophages, dendritic cells, granulocytes and mast cells. CD300a has a single Ig V-like extracellular domain which is homologous to the Ig-binding domains of the of the Fc receptor for polymeric IgA and IgM. CD300a is a non-MHC-specific inhibitory receptor that contains three immunoreceptor tyrosine-based inhibitory motifs in its intracellular region. CD300a can reportedly inhibit NK cell cytolytic activity and granulocyte and mast cell functions.
566342 Rev. 2
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
566342 Rev.2
Citations & References
Development References (7)
-
Alvarez Y, Tang X, Coligan JE, Borrego F. The CD300a (IRp60) inhibitory receptor is rapidly up-regulated on human neutrophils in response to inflammatory stimuli and modulates CD32a (FcgammaRIIa) mediated signaling.. Mol Immunol. 2008; 45(1):253-8. (Biology). 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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DeBell KE, Simhadri VR, Mariano JL, Borrego F. Functional requirements for inhibitory signal transmission by the immunomodulatory receptor CD300a.. BMC Immunol. 2012; 13:23. (Biology). View Reference
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Kim EJ, Lee SM, Suk K, Lee WH. CD300a and CD300f differentially regulate the MyD88 and TRIF-mediated TLR signalling pathways through activation of SHP-1 and/or SHP-2 in human monocytic cell lines.. Immunology. 2012; 135(3):226-35. (Clone-specific: Functional assay). View Reference
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Lankry D, Rovis TL, Jonjic S, Mandelboim O. The interaction between CD300a and phosphatidylserine inhibits tumor cell killing by NK cells.. Eur J Immunol. 2013; 43(8):2151-61. (Biology). View Reference
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Sabato V, Verweij MM, Bridts CH, et al. CD300a is expressed on human basophils and seems to inhibit IgE/FcεRI-dependent anaphylactic degranulation.. Cytometry B Clin Cytom. 2012; 82(3):132-8. (Clone-specific: Flow cytometry, Functional assay). View Reference
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
566342 Rev. 2
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.
