DESCRIPTION

OLR1 (also known as LOX1) is the receptor of oxidized low-density lipoprotein (ox-LDL) and is mainly expressed in endothelial cells, macrophages, smooth muscle cells, platelets, fibroblasts, and neurons. OLR1 is almost undetectable under physiological conditions, but it is upregulated following exposure to several proinflammatory and proatherogenic stimuli and can be detected in animal and human atherosclerotic lesions. OLR1 is the major contributor to ox-LDL uptake, internalization, and degradation in various cells. OLR1 plays a role in metabolism, cardiovascular disease, and cancer.

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	820.6	7.9	1.0
2	8	205.5	1.7	0.8
3	8	101.9	1.3	1.3

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	815.5	8.2	1.0
2	16	203.6	2.7	1.3
3	16	100.0	2.8	2.8

RECOVERY

The recovery of mouse OLR1/LOX1 spiked to three different levels throughout the range of the assay was evaluated.

Sample Type		Average% of Expected	Range (%)
Mouse serum	1:64	98	84-112
	1:128	90	88-94
Cell culture supernatant	1:32	89	87-90
	1:64	94	87-99

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 1:8. The cell culture supernatant was initially diluted 1:2.)

		Mouse serum	Cell culture supernatant
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	113	107
	Range (%)	105-121	100-115
1:8	Average% of Expected	123	117
	Range (%)	123-124	113-122
1:16	Average% of Expected	129	123
	Range (%)	128-130	117-130

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Mouse OLR1/LOX1 One-Step ELISA Kit produced at Sunlong Medical™.