Clone:
REA599
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
LPS R

Extended validation for CD14 Antibody, anti-human, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA599
MφP9+
M5E2++
61D3++
TÜK4++
63D3-
Cells were incubated with an excess of purified unconjugated CD14 (REA599) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for CD14. Human peripheral blood mononuclear cells (PBMCs) were stained with CD14 antibodies and with a suitable counterstaining. As a control, CD14 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD14. Human peripheral blood mononuclear cells (PBMCs) were stained with CD14 antibodies and with a suitable counterstaining. As a control, CD14 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD14. Human peripheral blood mononuclear cells (PBMCs) were stained with CD14 antibodies and with a suitable counterstaining. As a control, CD14 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD14. Human peripheral blood mononuclear cells (PBMCs) were stained with CD14 antibodies and with a suitable counterstaining. As a control, CD14 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD14. Human peripheral blood mononuclear cells (PBMCs) were stained with CD14 antibodies and with a suitable counterstaining. As a control, CD14 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD14. Human peripheral blood mononuclear cells (PBMCs) were stained with CD14 antibodies and with a suitable counterstaining. As a control, CD14 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD14. Human peripheral blood mononuclear cells (PBMCs) were stained with CD14 antibodies and with a suitable counterstaining. As a control, CD14 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for CD14. Human peripheral blood mononuclear cells (PBMCs) were stained with CD14 antibodies and with a suitable counterstaining. As a control, CD14 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using CD14 (REA599). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD14 (REA599). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using CD14 (REA599). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD14 Antibody, anti-human, REAfinity™

Overview

Clone REA599 recognizes the human CD14 antigen. CD14 is part of the functional heteromeric LPS receptor complex comprised of at least CD14, TLR4, and MD-2. It up-regulates cell surface molecules, including adhesion molecules. CD14 is strongly expressed on most human monocytes and macrophages in peripheral blood, other bodily fluids, and in various tissues such as lymph nodes and spleen. CD14 is weakly expressed on subpopulations of human neutrophils and myeloid dendritic cells.
Additional information: Clone REA599 displays negligible binding to Fc receptors.

Alternative names

LPS R

Detailed product information

Technical specifications

CloneREA599
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
Cross-reactivity
cynomolgus monkey (
Macaca fascicularis
)
, dog, goat, cat, rabbit, mink, bovine, pig, sheep
AntigenCD14
Alternative names of antigenLPS R
Molecular mass of antigen [kDa]37
Distribution of antigenmacrophages, monocytes, spleen
Entrez Gene ID929
RRIDAB_2655049, AB_2655050, AB_2655051, AB_2655052, AB_2655053, AB_2655054, AB_2655055, AB_2655056, AB_2655057, AB_2655058, AB_2655059, AB_2655060, AB_2655061, AB_2655062, AB_2655063, AB_2655064, AB_2655065, AB_2655066, AB_2655067, AB_2801871, AB_2655048

Resources for CD14 Antibody, anti-human, REAfinity™

References for CD14 Antibody, anti-human, REAfinity™

Publications

  1. Goyer, S. M. and Ferrero, E. (1987) Biochemical analysis of myeloid antigen and cDNA expression of gp55 (CD14). Oxford, Oxford University Press
  2. Wilson, A. D. et al. (1995)
    Selection of monoclonal antibodies for the identification of lymphocyte surface antigens in the New World primate
    Saguinus oedipus oedipus
    (cotton top tamarin).
    J. Immunol. Methods 178: 195-200
  3. Dzionek, A. et al. (2000) BDCA-2, BDCA-3, BDCA-4: Three markers for distinct subsets of dendritic cells in human peripheral blood. J. Immunol. 165: 6037-6046
  4. Neu, C. et al. (2013) CD14-dependent monocyte isolation enhances phagocytosis of listeria monocytogenes by proinflammatory, GM-CSF-derived macrophages. PLoS One 8(6): e66898
  5. Paccou, J. et al. (2013)
    Determination and modulation of total and surface calcium-sensing receptor expression in monocytes
    in vivo
    and
    in vitro
    .
    PLoS One 8(10): e74800

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