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BV421 Mouse Anti-Human CD14
BV421 Mouse Anti-Human CD14
Multiparameter flow cytometric analysis of CD14 expression on Human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon™ BV421 Mouse Anti-Human CD14 antibody (Cat. No. 567939/567940; Right Plot). 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 leucocytes. 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 BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon™ BV421 Mouse Anti-Human CD14 antibody (Cat. No. 567939/567940; Right Plot). 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 leucocytes. 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 Horizon™
monocyte differentiation antigen CD14; LPS receptor; LPS-R
Human (QC Testing)
Mouse IgG1, κ
Human Peripheral Blood Monocytes
Flow cytometry (Routinely Tested)
5 µl
929
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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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).
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
567939 Rev. 1
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.

567939 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
567939 Rev.1
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
567939 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.