DESCRIPTION

Surfeit locus protein 1 (SURF1) is a crucial component of the mitochondrial translation regulation assembly intermediate of cytochrome c oxidase complex (MITRAC complex). It plays a significant role in the biogenesis of cytochrome c oxidase (COX), a key enzyme in the mitochondrial electron transport chain responsible for oxidative phosphorylation. SURF1 aids in the correct assembly of COX by acting as a central assembly intermediate, receiving subunits imported to the inner mitochondrial membrane and regulating COX1 mRNA translation.

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 (pg/mL)	SD	CV%
1	8	1,698.5	76.6	4.5
2	8	673.8	52.4	7.8
3	8	396.7	21.5	5.4

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	1,556.6	158.3	10.2
2	16	681.8	93.9	13.8
3	16	400.6	19.3	4.8

RECOVERY

The recovery of human/rat SURF1 spiked to three different levels throughout the range of the assay in lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Lysate	1:8	114	107-126
	1:16	106	99-119

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 lysate was initially diluted 1:2. )

		Lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	110
	Range (%)	106-113
1:8	Average% of Expected	117
	Range (%)	107-126
1:16	Average% of Expected	101
	Range (%)	92-110

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human/rat SURF1 produced at Sunlong Medical™.