Human CD14 Magnetic Luminex® Performance Assay

Catalog #: LBAM383 Datasheet

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Quantist software graph of signal intensity versus concentration for CD14 in the Human Biomarker A Luminex Panel

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Summary Configure Your Assay Product Features Precision & Accuracy Data Examples Product Datasheets Background

Human CD14 Magnetic Luminex® Performance Assay Summary

Panel	Luminex Performance Human Biomarker Panel A
Assay Type	Magnetic bead-based multiplex assay for the Luminex^® platform
Format	1 x 96-well microplate and Magnetic antibody-coated beads
Species	Human
Analytes Detected	Please see analyte list in assay customization tool below.
Performance Validation	Our Luminex High Performance Assays undergo our most extensive validation testing. All Luminex High Performance Assay Panels are validated for use with serum and plasma samples. Some High Performance Assays are additionally validated for cell culture supernatants, milk, saliva, or urine, as indicated on the individual product data sheets. Luminex High Performance Assays are tested for sensitivity (three-quarters the low standard), intra-assay precision, inter-assay precision, and to ensure assay linearity for validated sample types. Recovery values for individual samples in validated sample types are also tested. Validation data for each analyte can be found in the product datasheet.

Make your selections below to configure your assay

Assays

Below are kits that are available based on your selections. These selections may be further customized before adding to the cart or emailing the configuration to your purchasing department or local distributor to place your order. Selected analytes may be included in more than one High Performance Assay. All assay options are displayed.

If you run into bead region incompatibilities, send your Luminex code to our team and find out more.

How would you like these analytes mixed?

Premixed

User Mixed

Please select at least one mixing option to see the kits.

Would you like to split based on dilution?

Do not split

Split based on serum

Split based on plasma

Split based on cell culture supernates

Not finding your analyte of interest? View our complete Luminex offering in the full version of our Luminex Assay Ordering Tool. We also offer Custom Luminex Services

Product Features

Magnetic Luminex High Performance Assays are flexible bead-based multiplex assays. They allow up to 50 user-defined target analytes to be simultaneously profiled using cell culture supernates, serum, or plasma samples. Some panels are also validated with human milk, saliva, and/or urine samples.

Simultaneously profile Human CD14 with up to analytes of your choice in one sample
Magnetic format allows for easier wash steps
Undergoes similar validation testing as Quantikine^® ELISA Kits
Precise and Specific
Requires a small sample volume (<50 µL)
Run your assay in just 3-5 hours
Mass-calibrated standards for consistent results with every new lot of material
Enough reagents for one 96-well plate

Base Kit Components

Standard Cocktail(s)
Bead Diluent
Biotin Antibody Diluent
Calibrator Diluent
Wash Buffer
Streptavidin-PE
One flat-bottom 96-well Microplate
Foil Plate Sealers (4)
Mixing Bottle
Standard Value Card**

**A standard curve must be generated each time an assay is run, utilizing values from the Standard Value Card included in the Base Kit.

Bead Set Contents

User-mix antibody-coated Magnetic beads in individual vials
User-mixed cocktail of biotinylated detection antibody

Precision

	Intra-Assay Precision			Inter-Assay Precision
Intra-assay Precision (precision within an assay) - Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (precision between assays) - Three samples of known concentration were tested in thirty-two separate assays to assess precision between assays.
Sample	1	2	3	1	2	3
n	20	20	20	32	32	32
Mean (pg/mL)	593	4427	25,835	591	4756	32,468
Standard deviation	33.2	172	2489	85.3	540	4287
CV (%)	5.6	3.9	9.6	14.4	11.3	13.2

Recovery

Sample Type					Average % Recovery	Range
Samples containing and/or spiked with human CD14 were evaluated for recovery.
Cell culture supernates					100	93-114%

Linearity

		Cell culture supernates*	Serum	EDTA plasma	Heparin plasma
Samples containing and/or spiked with human CD14 were serially diluted to evaluate assay linearity.
1:2	Average % of Expected	106	98	93	104
1:2	Range (%)	92-122	92-106	73-113	95-112
1:4	Average % of Expected	107	107	97	112
1:4	Range (%)	100-115	100-116	79-117	104-118
1:8	Average % of Expected	103	112	98	112
1:8	Range (%)	89-114	104-124	79-121	101-128

Which Luminex Assay is right for me?

Assays for the Luminex platform are offered as High Performance Assays or Assays. The Luminex High Performance Assays are fully validated panels with a focused selection of analytes. They are available in We Mix, You Mix, and Predetermined formats. The Luminex Assays allows for the maximum number of analytes in a multiplex and is supplied as a premixed kit. View a table comparing the features and benefits of these bead-based multiplex assays.

More Information About Magnetic Bead-based Multiplex Kits For The Luminex^® Platform

Magnetic Luminex High Performance Assays are designed for use with the Luminex MAGPIX CCD Imager. Alternatively, kits can be used with the Luminex 100/200™ or Bio-Rad^® Bio-Plex^®, dual laser, flow-based sorting and detection platforms.

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer. One laser is bead-specific and determines which analyte is being detected. A magnet in the analyzer captures and holds the superparaMagnetic microparticles in a monolayer. Two spectrally distinct Light Emitting Diodes (LEDs) illuminate the beads. One LED identifies the analyte that is being detected and the second LED determines the magnitude of the PE-derived signal, which is in direct proportion to the amount of analyte bound. Each well is imaged with a CCD camera. Kits can also be used with Luminex 100/200 or a Bio-Rad Bio-Plex dual laser, flow-based systems.

Scientific Data

N/A

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Quantist software graph of signal intensity versus concentration for CD14 in the Human Biomarker A Luminex Panel

Background: CD14

CD14 is an acute phase glycoprotein that binds lipopolysaccharide endotoxins (LPS) from Gram-negative bacteria. It is primarily expressed on monocytes, macrophages, and neutrophils, and soluble forms can be secreted or shed from the cell surface. CD14 accepts LPS from its initial binding by LBP, and it can further transfer LPS to the TLR4/MD2 complex. LPS acctivation of TLR4 triggers inflammatory activation. CD14 also contributes to the response to TLR3 ligands, phosphoinositides, as well as the surfactant proteins SP-A and SP-D.

Entrez Gene IDs

929 (Human); 12475 (Mouse); 60350 (Rat); 100037938 (Porcine)

Alternate Names

CD14 antigen; CD14 molecule; CD14; monocyte differentiation antigen CD14; Myeloid cell-specific leucine-rich glycoprotein