DESCRIPTION

Phospholipase A2 group IB, also know as PLA2G1B, is a secreted member of the phospholipase A2 (PLA2) class of enzymes, is produced by the pancreatic acinar cells. PLA2G1B is a calcium-dependent enzyme which can catalyzes the hydrolysis of the sn-2 position of membrane glycerophospholipids to release arachidonic acid (AA) and lysophospholipids. PLA2G1B is involved in several physiological processes including cell contraction, cell proliferation and pathological response.

PRINCIPLE OF THE ASSAY

A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody also binds to the analyte. Horseradish peroxidase (HRP)-conjugated secondary antibody binds to the detection antibody. 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	0.69	0.05	6.7
2	20	9.55	0.87	9.1
3	20	92.38	9.37	10.1

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	24	0.64	0.04	6.9
2	24	9.45	0.59	6.3
3	24	78.98	6.95	8.8

RECOVERY

The recovery of human PLA2G1B spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:2	101	72-115
	1:4	99	77-114
Cell culture supernatant	1:4	105	77-126

LINEARITY

To assess the linearity of the assay, three samples were spiked with high concentrations of human PLA2G1B in various matrices and diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. (The samples were initially diluted 1:2)

		Cell lysate	Cell culture supernatant
1:2	Average% of Expected	78	108
	Range (%)	76-80	107-110
1:4	Average% of Expected	85	83
	Range (%)	80-91	73-93
1:8	Average% of Expected	95	89
	Range (%)	83-107	82-95
1:16	Average% of Expected	99	90
	Range (%)	87-110	76-105

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human PLA2G1B produced at Sunlong Medical™.

Ordering & shipping

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