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Canine TNF-alpha Quantikine ELISA Kit

R&D Systems, part of Bio-Techne | Catalog # CATA00

R&D Systems, part of Bio-Techne
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CATA00

Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)

Sensitivity

4.2 pg/mL

Assay Range

7.8-500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)

Product Summary for Canine TNF-alpha Quantikine ELISA Kit

The Quantikine Canine TNF-alpha Immunoassay is a 4.5 hour solid-phase ELISA designed to measure canine TNF-alpha in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant canine TNF-alpha and antibodies raised against the recombinant protein. Results obtained for naturally occurring canine TNF-alpha showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural canine TNF-alpha.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Canine

Specificity

Natural and recombinant canine 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.

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 separate assays to assess inter-assay precision.

Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 100 66 106
Mean (pg/mL) 18.6 123 201 20.9 54.0 149
Standard Deviation 1.5 6.5 15.4 2.1 5.2 13.8
CV% 8.1 5.3 7.7 10.0 9.6 9.3

Recovery for Canine TNF-alpha Quantikine ELISA Kit

The recovery of canine TNF-alpha spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=7) 99 83-109
EDTA Plasma (n=6) 94 82-103
Heparin Plasma (n=6) 94 88-103
Serum (n=6) 92 83-106

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of canine TNF-alpha in each matrix were diluted with Calibrator Diluent and assayed.

Canine TNF-alpha ELISA Linearity

Scientific Data Images for Canine TNF-alpha Quantikine ELISA Kit

Canine TNF-alpha ELISA Standard Curve

Canine TNF-alpha ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. 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.

Background: TNF-alpha

TNF-alpha (Tumor necrosis factor alpha) plays a central role in inflammation, immune system development, apoptosis, and lipid metabolism. TNF-alpha was first identified as a cytotoxic factor produced by macrophages capable of killing mouse tumor cells. It is the prototypic ligand and along with Lymphotoxin-alpha, were identified as the first members of the TNF superfamily. Active TNF-alpha and other members of the TNF superfamily exist as a homotrimer with high structural homology. Receptor binding occurs at the interface of two TNF-alpha monomers. And receptor activation occurs when all three monomer interfaces are engaged with a receptor. For TNF-alpha, receptor binding and activation occurs through TNF R1 or TNF RII, and subsequently leads to activation of NF-kB or MAPK signaling pathways. Another pathway that TNF-alpha can activate utilizes the death domain of TNF RI to induce apoptosis. TNF-alpha promotes the inflammatory response largely through NF-kB signaling, and inhibition of TNF-alpha has proven successful in treating many autoimmune disorders. TNF-alpha is also present on the cell surface as membrane-bound TNF-alpha can induce the lysis of neighboring tumor cells and virus infected cells. TNF-alpha protein is translated as a type II transmembrane protein containing an N-terminal transmembrane domain. The soluble cytokine is released from its cell-anchoring TM domain by proteolytic processing by metalloproteases.

Long Name

Tumor Necrosis Factor alpha

Alternate Names

Cachetin, DIF, TNF, TNF-A, TNFA, TNFalpha, TNFG1F, TNFSF1A, TNFSF2

Entrez Gene IDs

7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)

Gene Symbol

TNF

Additional TNF-alpha Products

Product Documents for Canine TNF-alpha Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Canine TNF-alpha Quantikine ELISA Kit

For research use only

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⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.