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Multiplex Assay Kit for Alpha-Melanocyte Stimulating Hormone (aMSH) ,etc. by FLIA (Flow Luminescence Immunoassay)

α-MSH; Intermedins; Alpha-Melanotropin, Alpha-Melanocortin; Alpha-Intermedin

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 81-92 86
EDTA plasma(n=5) 91-101 98
heparin plasma(n=5) 93-101 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alpha-Melanocyte Stimulating Hormone (aMSH) ,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 Alpha-Melanocyte Stimulating Hormone (aMSH) ,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 Alpha-Melanocyte Stimulating Hormone (aMSH) ,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) 78-89% 87-94% 85-99% 81-97%
EDTA plasma(n=5) 78-98% 97-104% 98-105% 92-105%
heparin plasma(n=5) 79-94% 81-88% 90-101% 82-105%

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#:aMSH) 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 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
J Biochem. Generation of a human bone marrow-derived mesenchymal stem cell line expressing and secreting high levels of bioactive α-melanocyte-stimulating hormone. Pubmed: 23341471
Pigment Cell Melanoma Res Pharmacological induction of skin pigmentation unveils the neuroendocrine circuit regulated by light PubMed: 26582755
The Journal of Biological Chemistry Intermedin Restores Hyperhomocysteinemia-induced Macrophage Polarization and Improves Insulin Resistance in Mice Pubmed:27080257
Pigment Cell & Melanoma Research Pharmacological induction of skin pigmentation unveils the neuroendocrine circuit reCavia (Guinea pig )lated by light Pubmed:26582755
Nature Communications Endocrine disruptors induce perturbations in endoplasmic reticulum and mitochondria of human pluripotent stem cell derivatives. pubmed:28794470
Cell Stem Cell Super-Obese Patient-Derived iPSC Hypothalamic Neurons Exhibit Obesogenic Signatures and Hormone Responses Pubmed:29681516
European Neuropsychopharmacology Epigenetic alterations of the POMC promoter in tobacco dependence Pubmed:29871818
Neuropeptides Mechanisms of sustained long-term weight loss after RYGB: α-MSH is a key factor Pubmed:29685637
Neuropsychobiology Alcohol Withdrawal and Proopiomelanocortin Neuropeptides in an Animal Model of Alcohol Dependence Pubmed: 31117084
INTERNATIONAL IMMUNOPHARMACOLOGY Intermedin alleviates the inflammatory response and stabilizes the endothelial barrier in LPS-induced ARDS through the PI3K/Akt/eNOS signaling pathway Pubmed: 32892076
J Endocrinol Invest Serum alpha-melanocyte-stimulating hormone (a-MSH), brain-derived neurotrophic factor (BDNF), and agouti-related protein (AGRP) levels in children with Prader … Pubmed:35098494
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