DESCRIPTION

ROR1 is a transmembrane receptor that contains an extracellular section, a transmembrane segment, and a cytoplasmic region, a member of the RTK family of orphan receptors related to muscle-specific kinase and Trk neurotrophin receptors. ROR1 is primarily expressed by neural crest cells during embryogenesis. High expression of ROR1 is reported in many malignancies and is thought to be involved in tumor growth, apoptosis, and epithelial-mesenchymal transition. Moreover, ROR1 is also involved in the activation of signaling proteins important for cell proliferation, survival, and migration.

PRINCIPLE OF THE ASSAY

Sandwich ELISA structure (Detection antibody labeled with HRP) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with HRP also binds to the analyte. TMB acts as the HRP substrate and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns 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	19.43	0.80	4.14
2	8	4.25	0.29	6.73
3	8	2.17	0.13	5.94

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	19.16	1.22	6.34
2	16	4.41	0.30	6.82
3	16	2.22	0.15	6.74

RECOVERY

The recovery of human ROR1 spiked to three different levels throughout the range of the assay in cell lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:8	113	109-119
	1:16	117	114-120

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.

		Cell lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	94
	Range (%)	94-95
1:8	Average% of Expected	105
	Range (%)	99-109
1:16	Average% of Expected	121
	Range (%)	117-125

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human ROR1 Sandwich ELISA Kit produced at Sunlong Medical™.