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Mini Samples ELISA Kit for Transcription Factor P65 (NFKB3)

Product Specifications

Product Name Alternative

Nuclear factor NF-kappa-B p65 subunit; NFKBP65; RELA; p65; Nuclear Factor Of Kappa Light Polypeptide Gene Enhancer In B-Cells 3

Applications

Enzyme-linked immunosorbent assay for Antigen Detection.

Assay Principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Mini Samples Transcription Factor P65 (NFKB3) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Mini Samples Transcription Factor P65 (NFKB3) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Mini Samples Transcription Factor P65 (NFKB3), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Mini Samples Transcription Factor P65 (NFKB3) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Assay Protocol

1. Prepare all reagents, samples and standards; <br/> 2. Add 25&micro; L standard or sample to each well. Incubate 1 hour at 37&deg; C; <br/> 3. Aspirate and add 25&micro; L prepared Detection Reagent A. Incubate 1 hour at 37&deg; C; <br/> 4. Aspirate and wash 3 times; <br/> 5. Add 25&micro; L prepared Detection Reagent B. Incubate 30 minutes at 37&deg; C; <br/> 6. Aspirate and wash 5 times; <br/> 7. Add 25&micro; L Substrate Solution. Incubate 10-20 minutes at 37&deg; C; <br/> 8. Add 20&micro; L Stop Solution. Read at 450nm immediately.<br/>

Assay Performance Time

3H

Sample Type

Tissue homogenates, cell lysates and other biological fluids

Detection Range

0.156 - 10 ng/mL

Precision

<p> Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Mini Samples Transcription Factor P65 (NFKB3) were tested 20 times on one plate, respectively. <br/> Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Mini Samples Transcription Factor P65 (NFKB3) 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 0.058ng/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 Mini Samples Transcription Factor P65 (NFKB3) .<br/> No significant cross-reactivity or interference between Mini Samples Transcription Factor P65 (NFKB3) and analogues was observed. </p>

Method

Double-Antibody Sandwich

Organism Species

Homo sapiens (Human)

CAS Number

7732-18-5

Available Sizes

Curated Selection

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