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Multiplex Assay Kit for Midkine (MK) ,etc. by FLIA (Flow Luminescence Immunoassay)

MDK; ARAP; NEGF2; Neurite Growth-Promoting Factor 2; Amphiregulin-associated protein; Midgestation and kidney protein; Neurite outgrowth-promoting factor 2

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Midkine (MK) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Midkine (MK) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Midkine (MK) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Midkine (MK) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 88-103 95
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 93-102 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Midkine (MK) ,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 Midkine (MK) ,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 Midkine (MK) ,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) 78-92% 84-92% 89-102% 98-105%
EDTA plasma(n=5) 80-95% 91-104% 80-91% 89-97%
heparin plasma(n=5) 91-99% 88-95% 80-91% 98-105%

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#:MK) 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
Midkine: From Embryogenesis to Pathogenesis and Therapy Immunochemical Methods for the Detection of Midkine in Samples of Human Origin Springer: Source
33 Midkine Controls Arteriogenesis by Regulating the Bioavailability of Vascular Endothelial Growth Factor A and the Expression of Nitric Oxide Synthase 1 and … 10.1016:j.ebiom.2017.11.020
Circulation Journal Midkine Promotes Atherosclerotic Plaque Formation Through Its Pro-Inflammatory, Angiogenic and Anti-Apoptotic Functions in Apolipoprotein E-Knockout Mice pubmed:28781288
EBioMedicine Midkine Controls Arteriogenesis by Regulating the Bioavailability of Vascular Endothelial Growth Factor A and the Expression of Nitric Oxide Synthase 1 and … Pubmed:29233575
Journal of Clinical Laboratory Analysis Clinical value of jointly detection pleural fluid Midkine, pleural fluid adenosine deaminase, and pleural fluid carbohydrate antigen 125 in the identification of nonsmall … Pubmed:29797475
Charakterisierung der funktionalen Rolle des Zytokins Midkine bei der Arteriogenese :
Scientific Reports The possible role of Dickkopf-1, Golgi protein-73 and Midkine as predictors of hepatocarcinogenesis: a review and an Egyptian study Pubmed: 32198440
Odontology Effects of different pulp-capping materials on cell death signaling pathways of lipoteichoic acid-stimulated human dental pulp stem cells Pubmed: 33206337
Catalog No. Related products for research use of Rattus norvegicus (Rat) Organism species Applications (RESEARCH USE ONLY!)
RPA631Ra01 Recombinant Midkine (MK) Positive Control; Immunogen; SDS-PAGE; WB.
PAA631Ra01 Polyclonal Antibody to Midkine (MK) WB; IHC; ICC; IP.
LAA631Ra71 Biotin-Linked Polyclonal Antibody to Midkine (MK) WB; IHC; ICC.
MAA631Ra21 Monoclonal Antibody to Midkine (MK) WB; IHC; ICC; IP.
SEA631Ra ELISA Kit for Midkine (MK) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA631Ra Multiplex Assay Kit for Midkine (MK) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.