DESCRIPTION

Neutrophil gelatinase-associated lipocalin (NGAL) also known as Lipocalin-2 (LCN2), is a secreted glycoprotein that belongs to the lipocalin protein family. Several roles of NGAL were identified, including iron trafficking, chemotactic, and promoting cell differentiation and proliferation. NGAL was originally found in neutrophils and expressed in tissues containing renal, uterus, liver, spleen, cardiomyocytes, neurons, and immune cells. NGAL is a significant biomarker of cardiovascular diseases, ischemic stroke, acute renal injury, inflammation, and metabolic diseases.

PRINCIPLE OF THE ASSAY

An anti-tag antibody is pre-coated onto the bottom of wells. After adding antigen or samples, Tagged-Capture antibody and HRP-conjugated detection antibody, a sandwich complex is formed in the solution. TMB acts as a HRP substrate, and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns the 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	106.6	3.3	3.1
2	8	26.1	1.0	3.7
3	8	13.2	0.4	3.1

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	105.3	3.4	3.2
2	16	25.8	0.8	3.0
3	16	13.2	0.9	6.5

RECOVERY

The recovery of mouse Lipocalin-2/NGAL spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Mouse serum	1:1,000	98	96-100
	1:2,000	101	98-103
Cell culture supernatant	1:8,000	101	94-105
	1:16,000	97	96-101
Urine	1:1,000	101	100-103
	1:2,000	93	90-94

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 mouse serum was initially diluted 1:250. The cell culture supernatant was initially diluted 1:1,000. The urine was initially diluted 1:250.)

		Mouse serum	Cell culture supernatant	Urine
1:2	Average% of Expected	100	100	100
	Range (%)	-	-	-
1:4	Average% of Expected	102	99	101
	Range (%)	100-104	97-101	100-102
1:8	Average% of Expected	103	104	102
	Range (%)	101-105	103-104	102-103
1:16	Average% of Expected	106	106	104
	Range (%)	102-110	101-112	99-109

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Mouse Lipocalin-2/NGAL One-Step ELISA Kit produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
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