DESCRIPTION

Prolactin (PRL) is a pleiotropic hormone which is mainly synthetized by the lactotrophs of the anterior pituitary gland. Prolactin has a wide variety of effects. It causes the breasts to grow and make milk during pregnancy and after birth. Prolactin levels are normally high for pregnant women and new mothers. Prolactin also plays an essential role in metabolism, regulation of the immune system and pancreatic development. Normally during pregnancy prolactin serum levels are increased to hold of insulin resistance (IR). As well, prolactin is involved in regulation of glucose metabolism, since high prolactin levels are associated with low incidence of type 2 diabetes mellitus (T2DM).

PRINCIPLE OF THE ASSAY

Sandwich ELISA structure (Detection antibody labeled with biotin) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with biotin also binds to the analyte. Streptavidin-HRP binds to the biotin. 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 (pg/mL)	SD	CV%
1	20	101.7	8.5	8.4
2	20	305.6	5.6	1.8
3	20	1,054.2	21.8	2.1

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	24	72.9	4.1	5.1
2	24	264.1	7.1	2.7
3	24	958.0	28.2	2.9

RECOVERY

The recovery of human Prolactin 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	94	89-100
	1:16	103	102-110
Cell culture supernatant	1:2	101	95-109
	1:4	85	78-90
Human milk	1:60	102	94-107
	1:120	103	89-123

LINEARITY

To assess the linearity of the assay, cell culture supernatant samples were spiked with high concentrations of human Prolactin in various matrices and diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. Human serum and human milk were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay.(The human milk samples were initially diluted 1: 8 )

		Human serum(Sample Diluent PT 1-eg)	Cell culture supernatant(Sample Diluent PT 1-ag)	Human milk(Sample Diluent PT 3)
1:2	Average% of Expected	100	113	100
	Range (%)	-	96-124	-
1:4	Average% of Expected	100	106	104
	Range (%)	97-103	96-116	103-106
1:8	Average% of Expected	101	100	108
	Range (%)	97-107	91-112	107-111
1:16	Average% of Expected	109	99	111
	Range (%)	103-112	88-113	108-117

CALIBRATION

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

Ordering & shipping

• Minimum order: 1 kit
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• Price: USD, per kit (48T / 96T options available)