DESCRIPTION

Ubiquitin C-terminal hydrolase 1 (UCHL1), previously known as PGP9.5, is found primarily in neurons and neuroendocrine cells with a very high concentration in dendrites and perikarya. UCHL1 is a more recently recognized biomarker of neuronal injury. UCHL1 could be detected in the CSF and serum with a half-life of 7-9 hours after severe traumatic brain injury. Increased plasma concentrations of UCHL1 therefore reflect increased blood brain barrier permeability and destruction of neurons.

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 20 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 24 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	20	4.58	0.16	3.4
2	20	1.14	0.03	2.9
3	20	0.30	0.02	5.9

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	24	4.54	0.22	4.9
2	24	1.02	0.03	3.1
3	24	0.23	0.02	8.6

RECOVERY

The recovery of human UCH-L1 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human serum	1:8	89	84-98
	1:16	95	91-99
Cell lysate	1:800	108	92-123
	1:1600	99	72-127

LINEARITY

To assess the linearity of the assay, human serum, cell lysates were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. (The serum samples were initially diluted 1:2. The cell lysate samples were initially diluted 1:200. )

		Human serum(Sample Diluent PT 1B5)	Cell lysate(Sample Diluent PT 5)
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	81	90
	Range (%)	78-89	76-103
1:8	Average% of Expected	102	80
	Range (%)	96-110	74-90
1:16	Average% of Expected	102	82
	Range (%)	100-105	72-98

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human UCH-L1 produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
• Lead time: varies with order quantity and specifications. Please consult your local distributor or our sales team for confirmation
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• Price: USD, per kit (48T / 96T options available)