DESCRIPTION

Mouse PON3 (Paraoxonase 3) is a member of the paraoxonase gene family, possessing lactonase and arylesterase activities. It efficiently hydrolyzes substrates such as statin lactones (e.g., lovastatin) and N-acyl homoserine lactones. PON3 exhibits potent antioxidant activity and can inhibit low-density lipoprotein (LDL) oxidation. Gene knockout studies have shown that PON3deficient mice are prone to obesity, gallstone formation, and atherosclerosis. Furthermore, PON3 expression is upregulated in tumor tissues, where it promotes tumor progression by protecting mitochondria from oxidative stress and apoptosis. This protein is also involved in the regulation of the epithelial sodium channel (ENaC) and participates in sodium ion transport in the distal renal tubule. The expression level of PON3 is significantly higher in newborn mice than in adult mice, suggesting its important role during development.

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 (ng/mL)	SD	CV%
1	8	20.93	0.80	3.82
2	8	4.70	0.10	2.13
3	8	2.52	0.07	2.78

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	20.84	0.73	3.50
2	16	4.68	0.09	1.92
3	16	2.48	0.07	2.82

RECOVERY

The recovery of mouse PON3 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Mouse serum	1:16	121	116-128
	1:32	107	100-117
Lysate	1:8	91	86-93
	1:16	101	100-102

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 mouse serum was initially diluted 1:4. The lysate was initially diluted 1:100.)

		Mouse serum(Sample Diluent PT 5)	Lysate(Sample Diluent PT 3)
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	106	86
	Range (%)	104-109	79-94
1:8	Average% of Expected	110	88
	Range (%)	101-119	75-102
1:16	Average% of Expected	87	91
	Range (%)	80-94	77-104

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant mouse PON3 produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
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• Price: USD, per kit (48T / 96T options available)