DESCRIPTION

ACBP/DBI (acyl-CoA binding protein/Diazepam-binding inhibitor) is a widely expressed protein that binds long-chain fatty acylCoA esters and plays a role in fatty acyl-CoA transport and pool formation. ACBP/DBI is critical to cellular proliferation, differentiation, mitochondrial functions, and autophagy. Deletion of ACBP in mouse results in sebocyte hyperplasia and sparse, matted hair with a greasy appearance.

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	11.52	0.95	8.26
2	8	2.52	0.13	5.26
3	8	1.30	0.08	6.19

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	11.00	1.10	9.97
2	16	2.41	0.16	6.84
3	16	1.27	0.09	6.73

RECOVERY

The recovery of mouse ACBP/DBI 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	93	86-97
Tissue lysate	1:16	95	93-96

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 tissue lysate was initially diluted 1:16.)

		Mouse serum(Sample Diluent PT 3)	Tissue lysate(Sample Diluent PT 4B1)
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	100	103
	Range (%)	95-104	98-107
1:8	Average% of Expected	99	86
	Range (%)	90-108	83-89
1:16	Average% of Expected	105	77
	Range (%)	99-110	74-79

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Mouse ACBP/DBI Sandwich ELISA Kit produced at Sunlong Medical™.