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PE Rat Anti-Mouse CD368 (Clec4d)
PE Rat Anti-Mouse CD368 (Clec4d)
Multiparameter flow cytometric analysis of  CD368 (Clec4d) expression on BALB/c Mouse thioglycolate-elicited peritoneal exudate cells (Thio-PECs). BALB/c Mouse Thioglycolate-elicited peritoneal cells were stained with BD Pharmingen™ APC Rat Anti-Mouse Ly-6G antibody (Cat. No.560599) and with either PE Rat IgG1, κ Isotype Control (Cat. No. 553925; Left Plot) or PE Rat Anti-Mouse CD368 (Clec4d) antibody (Cat. No. 571181/571182; Right Plot) at 1.0 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of  CD368 (Clec4d) [or Ig Isotype control staining] versus Ly-6G was derived from gated events with the forward and side-light scatter characteristics of viable (DAPI-negative) peritoneal cell populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of  CD368 (Clec4d) expression on BALB/c Mouse thioglycolate-elicited peritoneal exudate cells (Thio-PECs). BALB/c Mouse Thioglycolate-elicited peritoneal cells were stained with BD Pharmingen™ APC Rat Anti-Mouse Ly-6G antibody (Cat. No.560599) and with either PE Rat IgG1, κ Isotype Control (Cat. No. 553925; Left Plot) or PE Rat Anti-Mouse CD368 (Clec4d) antibody (Cat. No. 571181/571182; Right Plot) at 1.0 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of  CD368 (Clec4d) [or Ig Isotype control staining] versus Ly-6G was derived from gated events with the forward and side-light scatter characteristics of viable (DAPI-negative) peritoneal cell populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
Dectin-3; macrophage C-type lectin; Mcl; mcl; Mpcl
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG1, κ
Mouse KLH-linked Peptides P160 and P134 of Mouse Clec4d EC Domain
Flow cytometry (Routinely Tested)
0.2 mg/ml
17474
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 BD® 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 BD® CompBeads to ensure that BD® CompBeads 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).
  7. For U.S. patents that may apply, see bd.com/patents.
571182 Rev. 1
Antibody Details
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3A4/Clec4d

The 3A4 monoclonal antibody (also known as, 3A4/Clec4d) specifically recognizes mouse CD368 which is also known as C-type lectin domain family 4 member D (Clec4d) and Macrophage C-type lectin (MCL). CD368 (Clec4d) is a single pass type II transmembrane glycoprotein that is encoded by Clec4d which belongs to the C-type lectin/C-type lectin-like domain (CTL/CTLD) superfamily. CD368 (Clec4d) is expressed by myeloid cells, including monocytes, macrophages, and neutrophils as well as by some dendritic cells. This calcium dependent lectin is involved in innate immune sensing of pathogen-associated molecular patterns (PAMPs) such as mycobacterial trehalose 6,6'-dimycolate. It functions as an endocytic receptor that may be involved in antigen clearance as well as antigen presentation to T cells including T-helper 1 (Th1)- and Th17-like T cells. The expression of CD368 (Clec4d) may be upregulated in response to various microbial stimuli including TLR agonists.

571182 Rev. 1
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
571182 Rev.1
Citations & References
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View product citations for antibody "571182" on CiteAb

Development References (3)

  1. Hupfer T, Schick J, Jozefowski K, Voehringer D, Ostrop J, Lang R. Stat6-Dependent Inhibition of Mincle Expression in Mouse and Human Antigen-Presenting Cells by the Th2 Cytokine IL-4.. Front Immunol. 2016; 7:423. (Clone-specific: Flow cytometry). View Reference
  2. Kerscher B, Dambuza IM, Christofi M, et al. Signalling through MyD88 drives surface expression of the mycobacterial receptors MCL (Clecsf8, Clec4d) and Mincle (Clec4e) following microbial stimulation.. Microbes Infect. 2016; 18(7-8):505-9. (Clone-specific: Flow cytometry). View Reference
  3. Kerscher B, Wilson GJ, Reid DM, et al. Mycobacterial receptor, Clec4d (CLECSF8, MCL), is coregulated with Mincle and upregulated on mouse myeloid cells following microbial challenge.. Eur J Immunol. 2016; 46(2):381-9. (Clone-specific: Flow cytometry). View Reference
571182 Rev. 1

 

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