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Invitrogen
CD14 Monoclonal Antibody (61D3), NovaFluor™ Blue 510, eBioscience™
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FIGURE: 1 / 3
CD14 Antibody (H019T03B01-A) in Flow
Normal human peripheral blood cells were unstained (left) or stained with CD14 Monoclonal Antibody, NovaFluor Blue 510 (right). All cells were co-stained with CD11b Monoclonal Antibody, eFluor 450 (Product # 48-0112-82). Total viable cells in the monocyte gate were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired on a 5-laser Cytek Aurora and unmixed with autofluorescence extraction.
Product Details
H019T03B01-A
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG1, kappa
Class
Monoclonal
Type
Antibody
Clone
61D3
Conjugate
Excitation/Emission Max
493/513 nm
View spectra
Form
Liquid
Concentration
0.6 µg/Test
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
Product Specific Information
Description
The 61D3 monoclonal antibody reacts with human CD14, a 53-55 kDa GPI-linked glycoprotein. CD14 is expressed on monocytes, interfollicular macrophages and some dendritic cells. Complexes of LPS and LBP (LPS-Binding Protein) bind with high affinity to monocytes through the surface CD14.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
The 61D3 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.6 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.
Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.
Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (for 25 µg or 25 test products, Cat. No. C001T02F01; for 100 µg or 100 test, Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 µL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 µL of CellBlox Plus Blocking Buffer per 100 µL of cell sample (10^3 to 10^8 cells).
NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Excitation: 493 nm; Emission: 513 nm; Laser: 488 nm (Blue) Laser
Target Information
CD14 is a 55 kDa GPI-anchored glycoprotein that is constitutively expressed on the surface of mature monocytes, macrophages, and neutrophils. CD14 also serves as a multifunctional lipopolysaccharide receptor, and is released to the serum both as a secreted and enzymatically cleaved GPI-anchored form. CD14 binds lipopolysaccharide molecule in a reaction catalyzed by lipopolysaccharide-binding protein (LBP), an acute phase serum protein. The soluble sCD14 can discriminate slight structural differences between lipopolysaccharides and is important for neutralization of serum allochthonous lipopolysaccharides by reconstituted lipoprotein particles. Further, CD14 has been shown to bind apoptotic cells, and can affect allergic, inflammatory and infectious processes. Alternative splicing results in multiple transcript variants encoding the same CD14 isoform. Diseases associated with CD14 dysfunction include mycobacterium chelonae infection and Croup.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
How to use the Panel Builder
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
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Bioinformatics
Protein Aliases: CD 14; CD14; cd14 monocyte; LPSR antibody; Monocyte differentiation antigen CD14; My23 antigen; Myeloid cell-specific leucine-rich glycoprotein; sCD14; soluble CD14
Gene Aliases: CD14
UniProt ID: (Human) P08571
Entrez Gene ID: (Human) 929
MyD88-dependent toll-like receptor signaling pathway
MyD88-independent toll-like receptor signaling pathway
receptor-mediated endocytosis
phagocytosis
apoptotic process
inflammatory response
cell surface receptor signaling pathway
I-kappaB kinase/NF-kappaB signaling
response to heat
lipopolysaccharide-mediated signaling pathway
response to magnesium ion
positive regulation of type I interferon production
positive regulation of interferon-gamma production
positive regulation of tumor necrosis factor production
negative regulation of MyD88-independent toll-like receptor signaling pathway
toll-like receptor 4 signaling pathway
response to tumor necrosis factor
TRIF-dependent toll-like receptor signaling pathway
toll-like receptor TLR1:TLR2 signaling pathway
toll-like receptor TLR6:TLR2 signaling pathway
innate immune response
response to ethanol
positive regulation of endocytosis
positive regulation of cytokine secretion
response to electrical stimulus
necroptotic process
cellular response to lipopolysaccharide
cellular response to lipoteichoic acid
cellular response to diacyl bacterial lipopeptide
cellular response to triacyl bacterial lipopeptide
apoptotic signaling pathway
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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