DESCRIPTION

Macrophage migration inhibitory factor (MIF) is an upstream regulator of the innate and adaptive immune response that is widely expressed and that exhibits chemokine-like activities. Altered MIF expression has been associated with a multitude of diseases ranging from inflammatory disorders like sepsis, lupus and rheumatoid arthritis to organ pathologies such as heart failure, myocardial infarction, acute kidney injury, organ fibrosis and a number of malignancies. In the cytoplasm, MIF is stored in vesiclelike structures and secreted in response to a number of stimuli including lipopolysaccharide (LPS), tumor necrosis factor (TNF)-α, hypoxia, H2O2, thrombin and angiotensin II. MIF regulates inflammatory processes by recruiting immune cells, amplifying cytokine production, inhibiting apoptosis and counter-regulating the immunosuppressive action of glucocorticoids.

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	3275.6	138.0	4.2
2	20	908.8	22.0	2.4
3	20	220.2	19.4	8.8

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	24	3173.4	99.9	3.1
2	24	866.7	29.0	3.3
3	24	175.9	18.8	10.7

RECOVERY

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

Sample Type		Average% of Expected	Range (%)
Human serum	1:40	106	87-119
	1:80	103	82-118
Urine	1:8	99	79-112
	1:16	113	86-127
Human milk	1:80	101	70-119
	1:160	103	72-126

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 and human milk samples were initially diluted 1:10. The urine samples were initially diluted 1:2. )

		Human serum (Samplediluent PT 3)	Urine (Sample diluent PT1)	Human milk (Samplediluent PT 3)
1:2	Average% of Expected	100	100	100
	Range (%)	-	-	-
1:4	Average% of Expected	122	110	103
	Range (%)	115-129	96-116	97-111
1:8	Average% of Expected	108	115	107
	Range (%)	97-119	96-128	96-113
1:16	Average% of Expected	116	99	105
	Range (%)	108-124	74-126	101-107

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human MIF 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
• Payment: T/T 100% in advance; PayPal; Alibaba Trade Assurance
• Shipping: FedEx / DHL / UPS express; cold-chain packing available on request
• Price: USD, per kit (48T / 96T options available)