CD14 Antibody, anti-human

Name: CD14 Antibody, anti-human
Availability: InStock

CD14 Antibody, anti-humanExtended Validation

Clone:

TÜK4

Type of antibody:

Primary antibodies

Isotype:

mouse IgG2aκ

Applications:

FC, MICS, IF, IHC, ICC

Alternative names:

LPS R

Flow cytometry applications forCD14 Antibody, anti-human

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770
PerCP
PerCP-Vio 700
VioBlue
VioGreen

Conjugate:

APC

Recommended dilution:

1:50

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Reported:

27467915, 27768888, 28174241, all publications

Products used:

130-113-705, 130-113-143

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD14 antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

MICS (MACSima Imaging Cyclic Staining) applications forCD14 Antibody, anti-human

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-113-705, 130-113-143

Compatible applications:

IF, IHC, ICC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-113-705, 130-113-143

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD14 Antibody, anti-human

Sensitivity

Performance comparison

Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.

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.

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.

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.

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

Comparison of staining pattern on non-fixed and fixed cells using CD14 (TÜK4). 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 (TÜK4). 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

Overview

Clone TÜK4 recognizes the human CD14 antigen and cross-reacts with non-human primate CD14. The CD14 antigen is a high affinity receptor for lipopolysaccharides (LPS) and LPS–binding protein (LBP)–complexes. It is part of the functional heteromeric LPS receptor complex comprised of CD14, TLR4, and MD-2.

CD14 is strongly expressed on most human monocytes and macrophages in peripheral blood, other body fluids, and various tissues, such as lymph nodes and spleen. CD14 is expressed at high levels, also on a few CD1c (BDCA-1)

⁺

CD2

⁺

myeloid dendritic cells and at low levels on neutrophilic granulocytes.

Ex vivo

differentiation of monocytes to dendritic cells is associated with down–regulation of CD14 antigen expression.

Alternative names

LPS R

Detailed product information

Technical specifications

Clone	TÜK4
Clonality	monoclonal
Isotype	mouse IgG2aκ
Isotype control	Isotype Control Antibody, mouse IgG2a
Host	mouse
Type of antibody	Primary antibodies
Species	human, non-human primate, other
Cross-reactivity	rhesus monkey ( Macaca mulatta ) , cynomolgus monkey ( Macaca fascicularis ) , pigtail monkey ( Macaca nemestrina ) , cotton-top tamarin ( Saguinus oedipus ) , pig, cow, sheep, goat, dog, mink, rabbit
Antigen	CD14
Alternative names of antigen	LPS R
Molecular mass of antigen [kDa]	37
Distribution of antigen	B cells, dendritic cells, epithelial cells, granulocytes, Langerhans cells, macrophages, monocytes, osteoclasts, platelets, plasma cells, liver
Entrez Gene ID	929
RRID	AB_2725974, AB_2726250, AB_2725975, AB_2726246, AB_2725971, AB_2726255, AB_2725980, AB_2726256, AB_2725981, AB_2726253, AB_2725978, AB_2726251, AB_2725976, AB_2726252, AB_2725977, AB_2726247, AB_2725972, AB_2726254, AB_2725979, AB_2726248, AB_2725973, AB_2726249

Resources for CD14 Antibody, anti-human

Documents and Protocols

App notes/customer reports

Isolation of monocytes with high purity directly from whole blood for transcriptome analysis in translational research.

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

Reviews for CD14 Antibody, anti-human

Staining Peripheral Blood with Anti-CD14 APC

1
2
3
4
5

CD14-APC, human (130-113-143)

Will continue to use this antibody.

Staining Peripheral Blood with Anti-CD14 APC

1
2
3
4
5

CD14-APC, human (130-113-705)

Will continue to use this antibody.

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References for CD14 Antibody, anti-human

Publications

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
Meissner, F. et al. (2010) Inflammasome activation in NADPH oxidase defective mononuclear phagocytes from patients with chronic granulomatous disease. Blood 116(9): 1570-1573
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
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
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
Jacobsen, C. N. et al. (1993) Reactivities of 20 anti-human monoclonal antibodies with leucocytes from ten different animal species. Vet. Immunol. Immunopathol. 39: 461-466
Goyer, S. M. and Ferrero, E. (1987) Biochemical analysis of myeloid antigen and cDNA expression of gp55 (CD14). Oxford, Oxford University Press

Applications

Extended validation