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PE Mouse Anti-Human CD14
PE Mouse Anti-Human CD14

Multiparameter flow cytometric analysis of CD14 expression on human peripheral blood leucocyte populations. Human whole blood was stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Human CD14 antibody (Cat. No. 567731/567732; Right Plot). After staining, the erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing the correlated expression of CD14 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Multiparameter flow cytometric analysis of CD14 expression on human peripheral blood leucocyte populations. Human whole blood was stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Human CD14 antibody (Cat. No. 567731/567732; Right Plot). After staining, the erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing the correlated expression of CD14 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Product Details
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BD Pharmingen™
monocyte differentiation antigen CD14; LPS receptor; LPS-R
Human (QC Testing)
Mouse IgG1, κ
Human Peripheral Blood Monocytes
Flow cytometry (Routinely Tested)
5 µl
Aqueous buffered solution containing BSA and ≤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 cells 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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  5. 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).
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Antibody Details
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63D3.rMAb

The 63D3.rMAb is a recombinant monoclonal antibody that specifically recognizes human CD14 which is also known as Monocyte differentiation antigen CD14, Myeloid cell-specific leucine-rich glycoprotein, or LPS Receptor (LPS-R). CD14 is expressed as an ~55 kDa glycophosphatidylinositol (GPI)-linked cell surface glycoprotein. CD14 is highly expressed on most mature monocytes and macrophages, and at lower levels on neutrophils. CD14 is also expressed on some dendritic cells (DC) and Langerhans cells. CD14 binds to bacterial cell wall components including lipopolysaccharide (LPS) bound to lipopolysaccharide-binding protein (LBP). CD14 serves as a co-receptor for certain signaling Toll-like receptors (TLRs) for the cellular detection and response to LPS as well as other pathogen-associated molecular patterns (PAMPs). Ligand-bound CD14 plays a role in stimulating various myeloid cells to produce pro-inflammatory cytokines and mediators and to upregulate the expression of cell surface adhesion molecules. Soluble forms of CD14 (sCD14) are reportedly released by cells into the blood or other bodily fluids. Soluble CD14 can regulate cellular responses to LPS and confer LPS-responsiveness to cells that do not ordinarily express CD14.

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 (4)

  1. Devitt A, Moffatt OD, Raykundalia C, Capra JD, Simmons DL, Gregory CD. Human CD14 mediates recognition and phagocytosis of apoptotic cells.. Nature. 1998; 392(6675):505-9. (Clone-specific: Blocking, ELISA, Functional assay, Western blot). View Reference
  2. Rosenberg SA, Ligler FS, Ugolini V, Lipsky PE. A monoclonal antibody that identifies human peripheral blood monocytes recognizes the accessory- cells required for mitogen-induced T lymphocyte proliferation.. J Immunol. 1981; 126(4):1473-7. (Immunogen: Bioassay, Flow cytometry, Fluorescence activated cell sorting). View Reference
  3. Ugolini V, Nunez G, Smith RG, Stastny P, Capra JD. Initial characterization of monoclonal antibodies against human monocytes.. Proc Natl Acad Sci USA. 1980; 77(11):6764-8. (Immunogen: Cytotoxicity, Flow cytometry, Radioimmunoassay). View Reference
  4. Wu Z, Zhang Z, Lei Z, Lei P. CD14: Biology and role in the pathogenesis of disease.. Cytokine Growth Factor Rev. 2019; 48:24-31. (Biology). View Reference
View All (4) 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.