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CLIA Kit for Tumor Necrosis Factor Alpha (TNFa)

Product Specifications

Product Name Alternative

DIF; TNF-A; TNFSF2; Cachectin; Tumor Necrosis Factor Ligand Superfamily Member 2

Applications

Chemiluminescent immunoassay for Antigen Detection.

Assay Principle

The microplate provided in this kit has been pre-coated with an antibody specific to Tumor Necrosis Factor Alpha (TNFa) . Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to Tumor Necrosis Factor Alpha (TNFa) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the Tumor Necrosis Factor Alpha (TNFa) level in the sample or standard.;

Assay Protocol

1. Prepare all reagents, samples and standards; <br/> 2. Add 100&micro; L standard or sample to each well. Incubate 2 hours at 37&deg; C; <br/> 3. Aspirate and add 100&micro; L prepared Detection Reagent A. Incubate 1 hour at 37&deg; C; <br/> 4. Aspirate and wash 3 times; <br/> 5. Add 100&micro; L prepared Detection Reagent B. Incubate 30 minutes at 37&deg; C; <br/> 6. Aspirate and wash 5 times; <br/> 7. Add 100&micro; L Substrate Solution. Incubate 10 minutes at 37&deg; C; <br/> 8. Read RLU value immediately.<br/>

Assay Performance Time

2H, 40min

Sample Type

Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids

Detection Range

1.37 - 1000 pg/mL

Precision

<p> Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Tumor Necrosis Factor Alpha (TNFa) were tested 20 times on one plate, respectively. <br/> Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Tumor Necrosis Factor Alpha (TNFa) were tested on 3 different plates, 8 replicates in each plate. <br/> CV (%) = SD/meanX100 <br/> Intra-Assay: CV<10% <br/> Inter-Assay: CV<12% <br/> </p>

Sensitivity

The minimum detectable dose of this kit is typically less than 0.50pg/mL

Stability

<p> The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. <br/> To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end. </p>

Specificity

<p> This assay has high sensitivity and excellent specificity for detection of Tumor Necrosis Factor Alpha (TNFa) .<br/> No significant cross-reactivity or interference between Tumor Necrosis Factor Alpha (TNFa) and analogues was observed. </p>

Method

Double-Antibody Sandwich

Organism Species

Rattus norvegicus (Rat)

Available Sizes

Curated Selection

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