Multiparameter flow cytometric analysis of CLEC5A (MDL-1) expression on human peripheral blood leucocytes populations. Human peripheral blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, incubated with Human BD Fc Block™ (Cat. No. 564219/564220), and then stained with either Alexa Fluor® 647 Mouse IgG2b, κ Isotype Control (Cat. No. 565378; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human CLEC5A (MDL-1) antibody (Cat. No. 566585; Right Plot) at 0.25 μg/test. Two-parameter flow cytometric contour plots showing the correlated expression of CLEC5A (MDL-1) [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 viable leucocyte populations. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System. A similar staining profile was observed when human blood was stained first followed by erythrocyte lysis with BD Pharm Lyse™ Lysing Buffer. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of CLEC5A (MDL-1) expression on human peripheral blood leucocytes populations. Human peripheral blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, incubated with Human BD Fc Block™ (Cat. No. 564219/564220), and then stained with either Alexa Fluor® 647 Mouse IgG2b, κ Isotype Control (Cat. No. 565378; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human CLEC5A (MDL-1) antibody (Cat. No. 566585; Right Plot) at 0.25 μg/test. Two-parameter flow cytometric contour plots showing the correlated expression of CLEC5A (MDL-1) [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 viable leucocyte populations. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System. A similar staining profile was observed when human blood was stained first followed by erythrocyte lysis with BD Pharm Lyse™ Lysing Buffer. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
CLEC5A; CLECSF5; MDL1; MDL-1; Myeloid DAP12-associating Lectin 1
Human (QC Testing)
Mouse IgG2b, κ
Human
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2869789
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
Human BD Fc Block™ RUO
Size
0.25 mg
Cat No.
564220
Human BD Fc Block™ RUO
Size
50 µg
Cat No.
564219
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Alexa Fluor® 647 Mouse IgG2b, κ Isotype Control RUO
Size
50 µg
Cat No.
565378
566585 Rev. 1
Antibody Details
283834
Monoclonal antibody 283834 specifically recognizes C-type lectin domain family 5, member A (CLEC5A) which is also known as C-type lectin superfamily member 5 (CLECSF5) or Myeloid DAP12-associating Lectin-1 (MDL-1). CLEC5A (MDL-1) is a ~40 kDa type II transmembrane glycoprotein that is expressed on immature myeloid cells, monocytes, macrophages, dendritic cells, and neutrophils. Clec5A (MDL-1) has a charged lysine in its transmembrane region that allows it to associate with DAP12 (aka, TYROBP) and transduce an activating signal in myeloid cells. Clec5A (MDL-1) regulates inflammatory responses during autoimmune arthritis and upon binding to Dengue viruses. The 283834 antibody does not crossreact with recombinant mouse Clec5a (MDL1) protein.
566585 Rev. 1
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
566585 Rev.1
Citations & References
Development References (5)
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Bakker AB, Baker E, Sutherland GR, Phillips JH, Lanier LL. Myeloid DAP12-associating lectin (MDL)-1 is a cell surface receptor involved in the activation of myeloid cells.. Proc Natl Acad Sci USA. 1999; 96(17):9792-6. (Biology). View Reference
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Chen ST, Liu RS, Wu MF, et al. CLEC5A regulates Japanese encephalitis virus-induced neuroinflammation and lethality.. PLoS Pathog. 2012; 8(4):e1002655. (Biology). View Reference
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González-Domínguez É, Domínguez-Soto Á, Nieto C, et al. Atypical Activin A and IL-10 Production Impairs Human CD16+ Monocyte Differentiation into Anti-Inflammatory Macrophages.. J Immunol. 2016; 196(3):1327-37. (Biology). View Reference
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Watson AA, Lebedev AA, Hall BA, et al. Structural flexibility of the macrophage dengue virus receptor CLEC5A: implications for ligand binding and signaling.. J Biol Chem. 2011; 286(27):24208-18. (Biology). View Reference
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Wortham BW, Eppert BL, Flury JL, et al. Cutting Edge: CLEC5A Mediates Macrophage Function and Chronic Obstructive Pulmonary Disease Pathologies.. J Immunol. 2016; 196(8):3227-31. (Biology). View Reference
566585 Rev. 1
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.
