DESCRIPTION

Progranulin, also known as Acrogranin, PCDGF, and Epithelin/Granulin Precursor, was first identified as a 593-aa secreted glycoprotein involved in the regulation of cancer progression and wound healing. It is a widely expressed glycoprotein that can undergo proteolysis to generate numerous Granulin peptides. Progranulin (PGRN) has been considered a growth factor-like molecule implicated in additional biological and pathological processes including early embryogenesis, inflammation, host defense, cartilage development and degradation.

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	57.29	1.61	2.8
2	20	102.27	4.25	4.2
3	20	189.93	7.59	4.0

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	24	56.16	2.84	5.1
2	24	96.77	5.2	5.4
3	24	182.62	13.12	7.2

RECOVERY

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

Sample Type		Average% of Expected	Range (%)
Human plasma	1:2	99	96-104
	1:4	100	92-110
Cell culture supernatant	1:2	113	100-124
	1:4	118	111-125
Human milk	1:4	111	84-127
	1:8	108	95-126
Urine	1:2	89	75-104
	1:4	101	74-124
Saliva	1:32	101	92-123
	1:64	103	99-113

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 human serum samples were initially diluted 1:2, the cell culture supernatant was initially diluted 1:4,the saliva were initially diluted 1:16, the human milk were initially diluted 1:16)

		Human serum(SampleDiluent PT 5-ef)	Cell culturesupernatant(Sample DiluentPT 5)	Urine (SampleDiluent PT 5-ef)	Human milk(Sample Diluent PT5-ef)	Saliva (SampleDiluent PT 5-ef)
1:2	Average% ofExpected	100	100	100	100	100
	Range (%)	-	-	-	-	-
1:4	Average% ofExpected	115	107	116	114	101
	Range (%)	110-121	103-111	109-122	102-124	78-128
1:8	Average% ofExpected	113	120	96	112	109
	Range (%)	103-123	117-125	81-110	96-125	94-117
1:16	Average% ofExpected		102		101	82
	Range (%)		99-105		88-124	70-94

CALIBRATION

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

Ordering & shipping

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