ELISA Kit for Dickkopf Related Protein 1 (DKK1)

SK; Dickkopf Homolog 1(Xenopus Laevis)

Specificity

This assay has high sensitivity and excellent specificity for detection of Dickkopf Related Protein 1 (DKK1).
No significant cross-reactivity or interference between Dickkopf Related Protein 1 (DKK1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Dickkopf Related Protein 1 (DKK1) and the recovery rates were calculated by comparing the measured value to the expected amount of Dickkopf Related Protein 1 (DKK1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-94 90
EDTA plasma(n=5) 87-94 91
heparin plasma(n=5) 78-102 84

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Dickkopf Related Protein 1 (DKK1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Dickkopf Related Protein 1 (DKK1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Dickkopf Related Protein 1 (DKK1) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 80-95% 82-101% 80-92% 92-105%
EDTA plasma(n=5) 83-104% 89-103% 89-96% 85-97%
heparin plasma(n=5) 98-105% 88-96% 78-99% 98-105%

Stability

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.
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.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Scientific Reports The possible role of Dickkopf-1, Golgi protein-73 and Midkine as predictors of hepatocarcinogenesis: a review and an Egyptian study Pubmed: 32198440
Nature Communications Immunological history governs human stem cell memory CD4 heterogeneity via the Wnt signaling pathway Pubmed: 32041953
JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY Circulating Osteocyte‐Related Biomarkers (vitamin D, sclerostin, dickkopf-1), hepcidin, and oxidative stress markers in early breast cancer: their impact in … Pubmed: 33065276
Oxid Med Cell Longev Long-Term Administration of Abacavir and Etravirine Impairs Semen Quality and Alters Redox System and Bone Metabolism in Growing Male Wistar Rats 34373766
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