Human Dkk-3 DuoSet ELISA Summary Assay Type | Solid Phase Sandwich ELISA | Format | 96-well strip plate | Assay Length | 4 hours | Sample Type & Volume Required Per Well | 100 μL | Sensitivity | 1.87 pg/mL | Assay Range | 31.2 - 2,000 pg/mL (Serum-free Cell Culture Supern, Mouse Serum, Rat Serum, Canine Serum, Porcine Serum) | Specificity | Natural and recombinant human Dkk-3 | 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. |
Product Summary This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Dkk-3. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet ELISA. Preparation and Storage Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. | Storage | Store the unopened product at 2 - 8 °C. Do not use past expiration date. |
Background: Dkk-3Members of the dickkopf-related protein family (Dkk-1, -2, -3, and -4) are secreted proteins with two cysteine-rich domains separated by a linker region. Dkk-3 and -4 also have one prokineticin domain. Dkk-1, -2, and -4 function as antagonists of canonical Wnt signaling by binding to LRP5/6, preventing interaction of LRP5/6 with Wnt-Frizzled complexes. Dkk-1, -2, and -4 also bind cell surface Kremen-1 or -2 and promote the internalization of LRP5/6. Antagonistic activity of Dkk-3 has not been demonstrated. Dkk proteins have distinct patterns of expression in adult and embryonic tissues and have a wide range of effects on tissue development and morphogenesis. The Dkk family also includes Soggy, which is homologous to Dkk-3 but not to the other family members. Soggy has not been shown to inhibit Wnt signaling, and its role in the pathway is unclear. Long Name: | Dickkopf-3 | Entrez Gene IDs: | 27122 (Human); 50781 (Mouse) | Alternate Names: | dickkopf (Xenopus laevis) homolog 3; dickkopf 3; dickkopf homolog 3 (Xenopus laevis); Dickkopf-3; dickkopf-related protein 3; Dkk3; Dkk-3; hDkk-3; regulated in glioma; REIC; REICdkk-3; RIG; RIG-like 5-6; RIG-like 7-1 |
Assay Procedure Refer to the product datasheet for the complete assay procedure. Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate. 1. Prepare all reagents, standard dilutions, and samples as directed in the product insert. 2. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal. 3. Add 50 μL of Assay Diluent to each well. 4. Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours. 5. Aspirate each well and wash, repeating the process 4 times for a total of 5 washes. 6. Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours. 7. Aspirate and wash 5 times. 8. Add 100 μL Substrate Solution to each well. 9. Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm. |
