ELISA Kit for Tyrosine Hydroxylase (TH) Homo sapiens (Human), Mus musculus (Mouse), Rattus norvegicus (Rat) Sandwich ELISA

TYH; DYT5b; Tyrosine 3-Monooxygenase; Dystonia 14

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  • ELISA Kit for Tyrosine Hydroxylase (TH) Packages (Simulation)
  • ELISA Kit for Tyrosine Hydroxylase (TH) Packages (Simulation)
  • ELISA Kit for Tyrosine Hydroxylase (TH) Results demonstration
  • SEB438Mi.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 81-91 85
EDTA plasma(n=5) 78-103 90
heparin plasma(n=5) 87-101 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Tyrosine Hydroxylase (TH) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Tyrosine Hydroxylase (TH) 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 Tyrosine Hydroxylase (TH) 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) 95-103% 94-102% 80-97% 82-93%
EDTA plasma(n=5) 86-101% 89-101% 86-94% 95-103%
heparin plasma(n=5) 95-103% 90-98% 81-102% 85-101%

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 1×120µL 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.

ELISA Kit for Tyrosine Hydroxylase (TH)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Tyrosine Hydroxylase (TH). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Tyrosine Hydroxylase (TH). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Tyrosine Hydroxylase (TH), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Tyrosine Hydroxylase (TH) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • 1, 25-dyhydroxyvitamin D3 Attenuates l-DOPA-Induced Neurotoxicity in Neural Stem CellsPubmed:25102940
  • Genetic Diagnosis of Two Dopa-Responsive Dystonia Families by Exome SequencingPubmed:25181484
  • Acetyl-l-carnitine protects dopaminergic nigrostriatal pathway in 6-hydroxydopamine-induced model of Parkinson's disease in the rat.pubmed:28199883
  • Elevated blood plasma levels of epinephrine, norepinephrine, tyrosine hydroxylase, TGFβ1, and TNFα associated with high-altitude pulmonary edema in an Indian population.pubmed:27540296
  • Silencing salusin-β attenuates cardiovascular remodeling and hypertension in spontaneously hypertensive rats.pubmed:28230187
  • Vaccarin administration ameliorates hypertension and cardiovascular remodeling in renovascular hypertensive ratspubmed:28681939
  • Multigenerational disruption of the thyroid endocrine system in marine medaka after a life-cycle exposure to perfluorobutanesulfonatePubmed:29565584
  • Cilostazol Mediated Nurr1 and Autophagy Enhancement: Neuroprotective Activity in Rat Rotenone PD ModelPubmed:29429051
  • Tetrabromobisphenol A caused neurodevelopmental toxicity via disrupting thyroid hormones in zebrafish larvaePubmed:29407805
  • The Effect of Recombinant Tyrosine Hydroxylase Expression on the Neurogenic Differentiation Potency of Mesenchymal Stem CellsPubmed:29656620
  • Montelukast attenuates rotenone-induced microglial activation/p38 MAPK expression in rats: Possible role of its antioxidant, anti-inflammatory and antiapoptotic effectsPubmed: 30222980
  • Silencing salusin β ameliorates heart failure in aged spontaneously hypertensive rats by ROS-relative MAPK/NF-κB pathways in the paraventricular nucleusPubmed: 30581101
  • Hydroxychloroquine antiparkinsonian potential: Nurr1 modulation versus autophagy inhibition
  • Perfluoropolyether carboxylic acids (novel alternatives to PFOA) impair zebrafish posterior swim bladder development via thyroid hormone disruptionPubmed: 31733528
  • Embryonic exposure to pentabromobenzene inhibited the inflation of posterior swim bladder in zebrafish larvaePubmed: 31935612
  • Nose to brain delivery of rotigotine loaded chitosan nanoparticles in human SH-SY5Y neuroblastoma cells and animal model of Parkinson's diseasePubmed: 32084576
  • Thyroid disruption and developmental toxicity caused by Cd2+ in Schizopygopsis younghusbandi larvaePubmed: 32344129
  • Selection of Cells for Parkinson's Disease Cell-Therapy
  • Effects of SiO2 nanoparticles on the uptake of tetrabromobisphenol A and its impact on the thyroid endocrine system in zebrafish larvae33385677
  • Tocotrienols protect differentiated SH-SY5Y human neuroblastoma cells against 6-hydroxydopamine-induced cytotoxicity by ameliorating dopamine biosynthesis and …Pubmed:35065349
  • Omarigliptin attenuates rotenone-induced Parkinson's disease in rats: Possible role of oxidative stress, endoplasmic reticulum stress and immune modulationPubmed:35439590
  • Unravelling the neuroprotective mechanisms of carotenes in differentiated human neural cells: biochemical and proteomic approachesPubmed:35415676
  • Parental and transgenerational impairments of thyroid endocrine system in zebrafish by 2, 4, 6-tribromophenol

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