Simple Plex Mouse TNF-alpha Cartridge

Catalog #: SPCKB-MP-000822 Datasheet
Simple Plex Mouse TNF-alpha assay kit for use on Ella instrument. Contains cartridge, sample diluent SD09, and wash buffer
Catalog # Availability Size / Price Qty
SPCKB-MP-000822
Control Products Available
Simple Plex Cartridge Kit for 16 samples, 1 analyte for use with Mouse Plasma / Serum.
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Simple Plex Mouse TNF-alpha Cartridge Summary

Assay Type
Automated sandwich ELISA
Format
Microfluidic cartridge containing three Glass Nano Reactors (GNR)
Assay Length
90 minutes
Sample Type & Volume Required Per Well*
Mouse Cell Culture Supernates (10 uL), Mouse Serum (25 uL), Mouse EDTA Plasma (25 uL), Mouse Heparin Plasma (25 uL)
Sensitivity
0.06 pg/mL
Assay Range
0.3 - 2,930 pg/mL (Mouse Cell Culture Supernates, Mouse Serum, Mouse EDTA Plasma, Mouse Heparin Plasma)
Specificity
Natural and recombinant mouse TNF-alpha.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

*The Sample Volume represented is based on the amount of sample incorporated into the reaction after taking into account the assay’s minimum required dilution for a given matrix. Serial dilution may be necessary to achieve some of the final sample volumes represented.

Product Summary

Simple Plex assay for the detection of mouse TNF-alpha in cell culture supernatant (CCS), serum, plasma (EDTA/Heparin).

Precision

Intra-Assay Precision (Precision within an assay) Each control was tested 16 times in one assay
Inter-Assay Precision (Precision between assays) Replicates of each control were tested in multiple assays performed by at least three technicians using two lots of reagents

Mouse Cell Culture Supernates, Mouse Serum, Mouse EDTA Plasma, Mouse Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 1 2
n 16 16 32 32
Mean (pg/mL) 8.38 391 8.3 427
Standard Deviation 0.748 38.1 1.08 53.4
CV% 8.9 9.8 13 12.5

Recovery

Recovery at three different spiked concentrations within the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Mouse EDTA Plasma (n=4) 101 97-103
Mouse Heparin Plasma (n=4) 102 93-109
Mouse Serum (n=4) 94 91-97

Linearity

Samples containing and/or spiked with high concentrations of mouse TNF-alpha were serially diluted with Sample Diluent to produce samples within the dynamic range of the assay
Mouse TNF-alpha  Ella Assay Linearity

Scientific Data

Mouse TNF-alpha Ella Assay Standard Curve

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Preparation and Storage

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

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.

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; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha

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