Multiplex Assay Kit for Myostatin (MSTN) ,etc. by FLIA (Flow Luminescence Immunoassay)

GDF8; Growth Differentiation Factor 8

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 79-95 87
EDTA plasma(n=5) 83-93 86
heparin plasma(n=5) 78-104 84
sodium citrate plasma(n=5) 83-102 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Myostatin (MSTN) ,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 Myostatin (MSTN) ,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 Myostatin (MSTN) ,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) 86-101% 82-97% 88-97% 79-97%
EDTA plasma(n=5) 93-102% 89-96% 88-101% 86-96%
heparin plasma(n=5) 78-92% 91-98% 91-105% 79-102%
sodium citrate plasma(n=5) 78-97% 83-99% 78-95% 92-101%

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#:MSTN) 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
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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