ELISA Kit for Alpha-Melanocyte Stimulating Hormone (aMSH)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 89-105 101
EDTA plasma(n=5) 92-99 95
heparin plasma(n=5) 78-94 84

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alpha-Melanocyte Stimulating Hormone (aMSH) 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) 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) 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) 80-94% 80-101% 95-105% 78-90%
EDTA plasma(n=5) 79-101% 93-102% 96-105% 79-90%
heparin plasma(n=5) 83-101% 78-89% 85-102% 96-103%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1 Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Reagent Diluent 1×300µL Stop Solution 1×6mL
TMB Substrate 1×9mL Instruction manual 1
Wash Buffer (30 × concentrate) 1×20mL

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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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
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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
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