DESCRIPTION

DKK3 (Dickkopf-3) is a secreted glycoprotein belonging to the Dickkopf family of Wnt signaling regulators. It is a multifaceted protein with complex, context-dependent roles in development, tissue homeostasis, and disease. DKK3 levels correlate with various disease states. It has been reported that DKK3 expression is decreased in many kinds of cancers, and it is thus thought to be a tumor suppressor gene. Low DKK3 is an independent risk factor for poor prognosis in pediatric acute lymphoblastic leukemia. Urinary DKK3 is being explored as a biomarker for kidney function decline.

PRINCIPLE OF THE ASSAY

An anti-tag antibody is pre-coated onto the bottom of wells. After adding antigen or samples, Tagged-Capture antibody and HRP-conjugated detection antibody, a sandwich complex is formed in the solution. TMB acts as a HRP substrate, and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns the solution yellow. The color intensity is proportional to the quantity of bound protein, which is measurable at 450 nm with the correction wavelength set at 630 nm.

PRECISION

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 8 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 16 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	8	2,623.6	47.8	1.8
2	8	656.1	21.5	3.3
3	8	323.3	13.6	4.2

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	2,579.2	74.5	2.9
2	16	638.0	25.2	3.9
3	16	314.4	16.7	5.3

RECOVERY

The recovery of human DKK3 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human serum	1:160	101	93-112
	1:320	104	89-111
Cell culture supernatant	1:32	95	70-126
	1:64	100	82-123

LINEARITY

To assess the linearity of the assay, samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. (The human serum was initially diluted 1:20. The cell culture supernatant was initially diluted 1:4.)

		Human serum	Cell culture supernatant
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	89	117
	Range (%)	84-94	109-125
1:8	Average% of Expected	82	119
	Range (%)	79-85	114-124
1:16	Average% of Expected	92	116
	Range (%)	92-93	103-129

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human DKK3 produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
• Lead time: varies with order quantity and specifications. Please consult your local distributor or our sales team for confirmation
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• Price: USD, per kit (48T / 96T options available)