ELISA Kit for Myostatin (MSTN) Rattus norvegicus (Rat) Competition ELISA

GDF8; Growth Differentiation Factor 8

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  • ELISA Kit for Myostatin (MSTN) Packages (Simulation)
  • ELISA Kit for Myostatin (MSTN) Packages (Simulation)
  • ELISA Kit for Myostatin (MSTN) Results demonstration
  • CEB653Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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) 81-98 90
EDTA plasma(n=5) 93-101 97
heparin plasma(n=5) 91-98 94

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) 91-101% 79-95% 80-103% 85-105%
EDTA plasma(n=5) 83-98% 92-101% 91-98% 97-105%
heparin plasma(n=5) 94-102% 96-105% 89-104% 91-101%

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
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.

ELISA Kit for Myostatin (MSTN)

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Myostatin (MSTN) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Myostatin (MSTN) and unlabeled Myostatin (MSTN) (Standards or samples) with the pre-coated antibody specific to Myostatin (MSTN). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Myostatin (MSTN) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Myostatin (MSTN) in the sample.

Citations

  • Anti-Myostatin Reduces Bone Mineral Loss in Ovariectomized RatsSapub:Source
  • Short-term, high-fat diet accelerates disuse atrophy and protein degradation in a muscle-specific manner in micePubMed: 26539241
  • Tumor inoculation site affects the development of cancer cachexia and muscle wastingPubMed: 26016447
  • Induction of transient tenogenic phenotype of high density cultured human dermal fibroblastsPubMed: 25748814
  • Higher Plasma Myostatin Levels in Cor Pulmonale Secondary to Chronic Obstructive Pulmonary DiseasePubmed:26998756
  • The Effect of Vibration Treatments Combined with Teriparatide or Strontium Ranelate on Bone Healing and Muscle in Ovariectomized RatsPubmed:27272029
  • Irisin and Myostatin Levels in Patients with Graves' Disease.pubmed:27986127
  • Effect of Strontium Ranelate on the Muscle and Vertebrae of Ovariectomized Ratspubmed:29242963
  • Compensatory anabolic signaling in the sarcopenia of experimental chronic arthritis.pubmed:28740214
  • Fermented corn-soybean meal elevated IGF1 levels in grower-finisher pigsPubmed: 30203098
  • Predictive value of serum myostatin for the severity and clinical outcome of heart failurePubmed: 31056368
  • Resistance training‐induced muscle hypertrophy is mediated by TGF‐β1‐Smad signaling pathway in male Wistar ratsPubmed: 31960436
  • Myostatin and muscle atrophy during chronic kidney diseasePubmed: 32974666
  • 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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