Human TNF-alpha Magnetic Luminex® Performance Assay

Name: Human TNF-alpha Magnetic Luminex® Performance Assay
Brand: R&D Systems
SKU: LUHM210
Rating: 5 (2 reviews)

R&D Systems, part of Bio-Techne | Catalog # LUHM210

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Citations (6)

COA / SDS

Luminex High Performance Assays

Luminex High Performance Assays go through rigorous development, validation, and quality control testing to ensure you are getting the best performance out of your multiplex assay. Panels are customizable so you can order a selection of the analytes or the entire panel.

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Key Product Details

Panel:	Luminex Performance Human Cytokine Panel A Luminex Performance Human Obesity Panel
Assay Type:	Magnetic bead-based multiplex assay for the Luminex^® MAGPIX^® platform
Format:	96-well microplate and antibody-coated magnetic beads*
Species:	Human
Analytes Detected:	TNF-alpha

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View Full Technical Details for Human TNF-alpha Magnetic Luminex® Performance Assay

Technical Data
Citations / Reviews (2)

Product Features for Human TNF-alpha Magnetic Luminex® Performance Assay

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 TNF-alpha with up to 21 analytes [Cytokine Panel A] or 9 analytes [Obesity Panel] 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

Performance Validation for Human TNF-alpha Magnetic Luminex® Performance Assay

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.

Kit Components for Human TNF-alpha Magnetic Luminex® Performance Assay

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 Certificate of Analysis provided with each Luminex Assay.

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 twenty-five separate assays to assess precision between assays.
Sample	1	2	3	1	2	3
n	20	20	20	25	25	25
Mean (pg/mL)	39.2	232	1326	44	234	1237
Standard deviation	1.66	8.56	63.9	3.0	15	87
CV (%)	4.2	3.7	4.8	7.3	6.2	7.0

Recovery

Sample Type	Average % Recovery	Range (%)
Samples containing and/or spiked with human TNF-a were evaluated for recovery.
Cell culture supernate	93	80-108
Serum	108	97-114
EDTA plasma	99	78-128
Heparin plasma	105	95-115

Linearity

		Cell culture supernate	Serum	EDTA plasma	Heparin plasma
Samples containing and/or spiked with human TNF-a were serially diluted to evaluate assay linearity.
1:2	Average % of Expected	108	104	107	101
1:2	Range (%)	96-117	96-116	95-122	91-108
1:4	Average % of Expected	108	107	110	98
1:4	Range (%)	90-121	96-123	90-135	94-105
1:8	Average % of Expected	108	110	111	96
1:8	Range (%)	87-120	87-120	67-145	96-98

Build Your Assay

Please choose one of the panels to display the corresponding analytes.

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

Which Luminex Assay is Right For Me?

Assays for the Luminex platform are offered as High Performance Assays or Discovery Assays . The Luminex High Performance Assays are fully validated panels with a focused selection of analytes. They are available as Configurable User Mix and Premixed options or as Fixed Panels. The Luminex Discovery Assays allow 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 on rndsystems.com .

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 200™ or FLEXMAP 3D, 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 a Luminex 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 FLEXMAP 3D dual laser, flow-based systems.

Scientific Data Examples for Human TNF-alpha Magnetic Luminex® Performance Assay

	Quantist software graph of signal intensity versus concentration for TNF-alpha in the Human Cytokine A Luminex Panel

Background: TNF-alpha

Tumor necrosis factor alpha (TNF-α), also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, apoptosis, and immune system development. TNF-α is produced by a wide variety of immune and epithelial cell types. Human TNF-α consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-α shares 97% aa sequence identity with rhesus and 71% - 92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-α. The 26 kDa type 2 transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer. Ligation of this complex induces reverse signaling that promotes lymphocyte costimulation but diminishes monocyte responsiveness.

Cleavage of membrane bound TNF-α by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-α. TNF-α trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers. TNF-α regulates lymphoid tissue development through control of apoptosis. It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages. TNF-α is a key cytokine in the development of several inflammatory disorders. It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism.

Item	Value
Long Name	Tumor Necrosis Factor alpha
Entrez Gene IDs	7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)
Alternate Names	APC1 protein, Cachectin, Cachetin, DIF, TNF, TNF, monocyte-derived, TNF-A, TNF-alpha, TNF-alphacachectin, TNFA, TNFATNF, macrophage-derived, TNFG1F, TNFSF1A, TNFSF2, TNFSF2TNF superfamily, member 2, TNFalpha, Tumor necrosis factor ligand superfamily member 2, tumor necrosis factor, tumor necrosis factor (TNF superfamily, member 2), tumor necrosis factor alpha, tumor necrosis factor-alpha

Preparation and Storage

Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage	Store the unopened product at 2 - 8 °C. Do not use past expiration date.