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Multiplex Assay Kit for Milk Fat Globule EGF Factor 8 (MFGE8) ,etc. by FLIA (Flow Luminescence Immunoassay)

HMFG; MFGM; SED1; BA46; EDIL1; OAcGD3S; SPAG10; hP47; Medin; Lactadherin; Sperm Associated Antigen 10; Sperm Surface Protein hP47; Milk fat globule-EGF factor 8

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Milk Fat Globule EGF Factor 8 (MFGE8) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Milk Fat Globule EGF Factor 8 (MFGE8) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Milk Fat Globule EGF Factor 8 (MFGE8) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Milk Fat Globule EGF Factor 8 (MFGE8) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-92 87
EDTA plasma(n=5) 79-94 87
heparin plasma(n=5) 89-97 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Milk Fat Globule EGF Factor 8 (MFGE8) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Milk Fat Globule EGF Factor 8 (MFGE8) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Milk Fat Globule EGF Factor 8 (MFGE8) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 91-103% 79-97% 81-88% 87-101%
EDTA plasma(n=5) 95-102% 83-93% 80-96% 90-104%
heparin plasma(n=5) 89-98% 97-104% 89-104% 88-99%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:MFGE8) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Fertility and Sterility Tumor necrosis factor α up-regulates endometrial milk fat globule-epidermal growth factor 8 protein production via nuclear factor κB activation, resulting in cell migration of epithelial cells. Pubmed: 24262600
U.S. Patent Application Biomarker for amyloid-b-related neurological disorders US20130302838
IUBMB Life. Elevated serum milk fat globule-epidermal growth factor 8 levels in type 2 diabetic patients are suppressed by overweight or obese status. pubmed:28067023
Journal of diabetes investigation Circulating milk fat globule-epidermal growth factor 8 levels are increased in pregnancy and gestational diabetes mellitus. pubmed:28035763
The Journal of Steroid Biochemistry and Molecular Biology Modifications of Western-type diet regarding protein, fat and sucrose levels as modulators of steroid metabolism and activity in liver pubmed:27471150
Journal of Steroid Biochemistry and Molecular Biology Depressive-like effect of prenatal exposure to DDT involves global DNA hypomethylation and impairment of GPER1/ESR1 protein levels but not ESR2 and AHR/ARNT signaling pubmed:28263910
Oncotarget Pantoprazole blocks the JAK2/STAT3 pathway to alleviate skeletal muscle wasting in cancer cachexia by inhibiting inflammatory response. pubmed:28489606
Molecular and Cellular Neuroscience TGF-β1 enhances phagocytic removal of neuron debris and neuronal survival by olfactory ensheathing cells via integrin/MFG-E8 signaling pathway 10.1016/j.mcn.2017.08.006
Amyloid Needle-shaped amyloid deposition in rat mammary gland: evidence of a novel amyloid fibril protein Pubmed: 31615282
clinical and translational medicine Milk fat globule EGF factor 8 restores mitochondrial function via integrin©\medicated activation of the FAK©\STAT3 signaling pathway in acute pancreatitis 33634976
Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
RPB286Mu02 Recombinant Milk Fat Globule EGF Factor 8 (MFGE8) Positive Control; Immunogen; SDS-PAGE; WB.
RPB286Mu01 Recombinant Milk Fat Globule EGF Factor 8 (MFGE8) Positive Control; Immunogen; SDS-PAGE; WB.
PAB286Mu01 Polyclonal Antibody to Milk Fat Globule EGF Factor 8 (MFGE8) WB; IHC; ICC; IP.
PAB286Mu02 Polyclonal Antibody to Milk Fat Globule EGF Factor 8 (MFGE8) WB; IHC; ICC; IP.
SEB286Mu ELISA Kit for Milk Fat Globule EGF Factor 8 (MFGE8) Enzyme-linked immunosorbent assay for Antigen Detection.
LMB286Mu Multiplex Assay Kit for Milk Fat Globule EGF Factor 8 (MFGE8) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.