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|Antigen||Dickkopf Homolog 1 (Xenopus Laevis) (DKK1) ELISA Kits|
Kits with alternative reactivity to:
|Method Type||Sandwich ELISA|
|Detection Range||100-30000 pg/mL|
|Minimum Detection Limit||100 pg/mL|
|2 references available|
|Supplier||Log in to see|
Product Details DKK1 ELISA KitTarget details Application Details Handling ProductDetails: References for DKK1 Kit (ABIN624961) Images
|Purpose||Human DKK-1 ELISA Kit for cell culture supernatants, plasma, and serum samples.|
|Sample Type||Plasma, Cell Culture Supernatant, Serum|
|Specificity||This ELISA kit shows no cross-reactivity with the following cytokines tested: human Angiogenin, BDNF, BLC, ENA-78, FGF- 4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin (OB), MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.|
|Sensitivity||< 100 pg/mL|
|Material not included||
Target detailsProduct Details DKK1 ELISA Kit Application Details Handling ProductDetails: References for DKK1 Kit (ABIN624961) Images back to top
|Alternative Name||Dkk-1 (DKK1 ELISA Kit Abstract)|
|Background||The Human Dkk-1 (Dickkopf homolog 1) ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human Dkk-1 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human Dkk-1 coated on a 96-well plate. Standards and samples are pipetted into the wells and Dkk-1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human Dkk-1 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Dkk-1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.|
|Pathways||WNT Signaling, Regulation of Muscle Cell Differentiation, Positive Regulation of fat Cell Differentiation|
Application DetailsProduct Details DKK1 ELISA Kit Target details Handling ProductDetails: References for DKK1 Kit (ABIN624961) Images back to top
|Application Notes||Recommended Dilution for serum and plasma samples2 fold|
|Sample Volume||100 μL|
1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
2. Sample dilution: 1x Assay Diluent D (Item K) should be used for dilution of serum/plasma samples. 1x Assay Diluent B should be used for dilution of cell culture supernates/urine. Suggested dilution for normal serum/plasma: 2 fold*. *Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
3. Assay Diluent B and Assay Diluent D should be diluted 5-fold with deionized or distilled water before use.
4. Preparation of standard: Briefly spin the vial of Item C and then add 1000 µL 1x Assay Diluent D (for serum/plasma samples) or 1x Assay Diluent B (for cell culture supernates/urine) into Item C vial to prepare a 900 ng/mL standard solution. Dissolve the powder thoroughly by a gentle mix. Add 20 µL 300 ng/mL standard from the vial of Item C, into a tube with 580 µL 1x Assay Diluent D or Assay Diluent B to prepare a 30,000 pg/mL standard solution. Pipette 300 µL 1x Assay Diluent D or 1x Assay Diluent B into each tube. Use the 30,000 pg/mL standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Assay Diluent D or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 300 µL 20 µL standard + 580 µL 200myl 200 µL 200 µL 200 µL 200 µL 200 µL 30,000 12,000 4,800 1,920 768 307.2 122.9 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent D (for serum/plasma samples) or 1x Assay Diluent B (cell culture medium/urine) into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent D (for serum/plasma samples) or 1x Assay Diluent B (cell culture medium/urine) and used in step 4 of Part VI Assay Procedure.
7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) ) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 700-fold with 1x Assay Diluent B (cell culture medium/urine or 1x Assay Diluent D (for serum/plasma samples). For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 20 µL of HRP-Streptavidin concentrate into a tube with 14 ml 1x Assay Diluent D (for serum/plasma samples) or 1x Assay Diluent B (cell culture medium/urine) to prepare a final 700 fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
5. Discard the solution. Repeat the wash as in step
6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
7. Discard the solution. Repeat the wash as in step
8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
|Calculation of Results||
Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Human Dkk-1 concentration (pg/mL) O D =4 50 n m 0.01 0.1 1 10 Assay Diluent D 10 100 1,000 10,000 100,000 Human Dkk-1 concentration (pg/mL) O D =4 50 n m 0.01 0.1 1 10 Assay Diluent B 10 100 1,000 10,000 100,000
Sensitivity: The minimum detectable dose of Dkk-1 is typically less than 100 pg/mL.
Recovery: Recovery was determined by spiking various levels of human Dkk-1 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 92.3 62-130 Plasma 72.8 59-110 Cell culture media 108.3 93-123
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 98.1 95.6 99.5 Range ( %) 89-112 84-103 90-113 1:4 Average % of Expected 96.2 92.5 96.3 Range ( %) 88-111 80-102 87-109
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %
|Assay Precision||Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %|
|Restrictions||For Research Use only|
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|Handling Advice||Avoid repeated freeze-thaw cycles.|
|Storage Comment||The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.|
|Expiry Date||6 months|
ProductDetails: References for DKK1 Kit (ABIN624961)Product Details DKK1 ELISA Kit Target details Application Details Handling Images back to top
|Product cited in:||
de Andrade, de Castro, Vicente, da Rosa, Nader, Pereira, Fröde: "Evaluation of circulating levels of inflammatory and bone formation markers in axial spondyloarthritis." in: International immunopharmacology, Vol. 21, Issue 2, pp. 481-6, 2014
Li, Huang, Tong, Wang, Zhang, Wang, Dai, Li, Lin, Wu: "Comparison of the regulation of ?-catenin signaling by type I, type II and type III interferons in hepatocellular carcinoma cells." in: PLoS ONE, Vol. 7, Issue 10, pp. e47040, 2012
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