ELISA Kit for Myostatin (MSTN)

GDF8; Growth Differentiation Factor 8

Specificity

This assay has high sensitivity and excellent specificity for detection of Myostatin (MSTN).
No significant cross-reactivity or interference between Myostatin (MSTN) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Myostatin (MSTN) and the recovery rates were calculated by comparing the measured value to the expected amount of Myostatin (MSTN) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-103 88
EDTA plasma(n=5) 89-96 93
heparin plasma(n=5) 92-99 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Myostatin (MSTN) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Myostatin (MSTN) 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 Myostatin (MSTN) 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) 81-96% 91-101% 81-96% 97-105%
EDTA plasma(n=5) 84-101% 87-103% 91-98% 80-101%
heparin plasma(n=5) 85-99% 91-105% 78-101% 92-105%

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
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 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

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

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Magazine Citations
Clinical Medicine and Diagnostics Anti-Myostatin Reduces Bone Mineral Loss in Ovariectomized Rats Sapub:Source
Nutr Metab (Lond) Short-term, high-fat diet accelerates disuse atrophy and protein degradation in a muscle-specific manner in mice PubMed: 26539241
International Journal of Cancer Tumor inoculation site affects the development of cancer cachexia and muscle wasting PubMed: 26016447
Connect Tissue Res Induction of transient tenogenic phenotype of high density cultured human dermal fibroblasts PubMed: 25748814
PLoS One Higher Plasma Myostatin Levels in Cor Pulmonale Secondary to Chronic Obstructive Pulmonary Disease Pubmed:26998756
Calcif Tissue Int. The Effect of Vibration Treatments Combined with Teriparatide or Strontium Ranelate on Bone Healing and Muscle in Ovariectomized Rats Pubmed:27272029
Archives of medical research Irisin and Myostatin Levels in Patients with Graves' Disease. pubmed:27986127
Calcified Tissue International Effect of Strontium Ranelate on the Muscle and Vertebrae of Ovariectomized Rats pubmed:29242963
Scientific Reports Compensatory anabolic signaling in the sarcopenia of experimental chronic arthritis. pubmed:28740214
Journal of Animal Science Fermented corn-soybean meal elevated IGF1 levels in grower-finisher pigs Pubmed: 30203098
European Journal of Internal Medicine Predictive value of serum myostatin for the severity and clinical outcome of heart failure Pubmed: 31056368
J Cell Physiol Resistance training‐induced muscle hypertrophy is mediated by TGF‐β1‐Smad signaling pathway in male Wistar rats Pubmed: 31960436
NEPHROLOGY DIALYSIS TRANSPLANTATION Myostatin and muscle atrophy during chronic kidney disease Pubmed: 32974666
Oxid Med Cell Longev Gender-Specific Behaviour in Obesity Stages I-II: Imbalance of Aminothiol Status and Adipomyokine Profile in Subjects with Different Insulin Resistance … 34868459
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