ELISA Kit for Thyrotropin Releasing Hormone (TRH)

TRF; Thyrotropin-Releasing Factor; Thyroliberin; Protirelin; Pro-tirelin; Prothyroliberin; TSH-releasing factor

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 81-103 91
EDTA plasma(n=5) 90-97 94
heparin plasma(n=5) 95-104 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Thyrotropin Releasing Hormone (TRH) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Thyrotropin Releasing Hormone (TRH) 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 Thyrotropin Releasing Hormone (TRH) 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) 79-101% 97-105% 94-101% 84-98%
EDTA plasma(n=5) 85-102% 79-102% 82-93% 81-104%
heparin plasma(n=5) 84-104% 78-101% 80-89% 81-104%

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 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
Plos one Thyrotropin-Releasing Hormone (TRH) Promotes Wound Re-Epithelialisation in Frog and Human Skin PubMed: PMC3759422
Analyst Ultrasensitive detection of thyrotropin-releasing hormone based on azo coupling and surface-enhanced resonance Raman spectroscopy Pubmed:27338554
journal of neuroendocrinology Thyroid dysfunction in children with autism spectrum disorder is associated with folate receptor α autoimmune disorder. pubmed:28199771
Journal of Neuroendocrinology Thyroid dysfunction in children with autism spectrum disorder is associated with folate receptor α autoimmune disorder 10.1111/jne.12461
Frontiers in Molecular Neuroscience Impact of Triclosan on Female Reproduction through Reducing Thyroid Hormones to Suppress Hypothalamic Kisspeptin Neurons in Mice Pubmed:29403355
Cell Reports Zika Virus Infection in Hypothalamus Causes Hormone Deficiencies and Leads to Irreversible Growth Delay and Memory Impairment in Mice Pubmed: 30404008
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