Rat Tyrosine Hydroxylase(TH) ELISA Kit

Code CSB-E13102r
Size 96T,5×96T,10×96T
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Product Details

Target Name
tyrosine hydroxylase
Alternative Names
Th ELISA Kit; Tyrosine 3-monooxygenase ELISA Kit; EC 1.14.16.2 ELISA Kit; Tyrosine 3-hydroxylase ELISA Kit; TH ELISA Kit
Abbreviation
TH
Uniprot No.
Species
Rattus norvegicus (Rat)
Sample Types
serum, tissue homogenates
Detection Range
78 pg/mL-5000 pg/mL
Sensitivity
19.5 pg/mL
Assay Time
1-5h
Sample Volume
50-100ul
Detection Wavelength
450 nm
Research Area
Metabolism
Assay Principle
quantitative
Measurement
Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<10%      
Three samples of known concentration were tested in twenty assays to assess.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of rat TH in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:100 Average % 90  
Range % 85-96  
1:200 Average % 96  
Range % 91-105  
1:400 Average % 98  
Range % 92-104  
1:800 Average % 94  
Range % 86-98  
Recovery
The recovery of rat TH spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range  
Serum (n=5) 90 85-94  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
pg/ml OD1 OD2 Average Corrected  
5000 2.563 2.501 2.532 2.390  
2500 2.113 2.131 2.122 1.980  
1250 1.683 1.699 1.691 1.549  
625 1.142 1.168 1.155 1.013  
312 0.712 0.754 0.733 0.591  
156 0.418 0.431 0.425 0.283  
78 0.322 0.341 0.332 0.190  
0 0.141 0.143 0.142    
Troubleshooting
and FAQs
Storage
Store at 2-8°C. Please refer to protocol.
Lead Time
3-5 working days after you place the order, and it takes another 3-5 days for delivery via DHL or FedEx
Description

This Rat tyrosine hydroxylase (TH) ELISA Kit was designed for the quantitative measurement of Rat tyrosine hydroxylase (TH) protein in serum, tissue homogenates. It is a Sandwich ELISA kit, its detection range is 78 pg/mL-5000 pg/mL and the sensitivity is 19.5 pg/mL .

Customer Reviews and Q&A

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 Q&A
Q:

I would like to know if fixed 4% PFA tissue will work in this kit?

A:
Thanks for your inquiry.
PFA you mentioned should be paraformaldehyde. This reagent will cause protein denaturation and usually it is used for immunohistochemistry. It is not suitable to elisa. If you want to test tissue samples, we suggest you follow our sample preparation method.
Tissue Homogenates 100mg tissue was rinsed with 1X PBS, homogenized in 1 ml of 1X PBS and stored overnight at -20°C. After two freeze-thaw cycles were performed to break the cell membranes, the homogenates were centrifuged for 5 minutes at 5000 x g, 2 - 8°C. The supernate was removed and assayed immediately. Alternatively, aliquot and store samples at -20°C or -80°C. Centrifuge the sample again after thawing before the assay. Avoid repeated freeze-thaw cycles. Pls let me know if you have any further questions. Thank you.
Q:

I would like to know if can use another lysis buffer with the PBS for homogenization? So he can protect the protein from degradation. Or should he just stick to using PBS only?

A:
Thanks for your inquiry.
CSB-E13102r
If you want to test tissue samples, we suggest you use PBS repeated freezing and thawing method.
If you use another lysis buffer with the PBS for homogenization, you need to make sure the lysis buffer doesn't contain any component causing protein degeneration. Some data shows that low concentration of NP、Tween-20 will not affect elisa too much, but the specific concentration range has not been validated. If your sample has been prepared, you can have a try, but we are not sure about the test result. We still suggest you use PBS method.
Pls let me know if you have any further questions. Thank you.
Q:

I am planning to run trail run with the plate and is not using the whole plate wells.
Is it possible if he had it in separate columns to run it?
Are there two vials of standard provided in the kit, so they can assay twice? And the stock solution of the standard can be stored at 2-8℃ for 5 days?

A:
Thanks for your inquiry.
The standard in the kit is lyophilized and there are 2 vials of standard in this kit.
Centrifugal resuspension is required before use. The standard solution after heavy suspension can only be stored for 2-8 degrees to avoid contamination and should be used up within a week. Pls let me know if you have any further questions. Thank you.
Q:

Thank you for your help and patience.I have another question about the kit. As he states in his own words:
“I just got the last question for you, the adhesive strip, If we are using the portion of the wells, can this be cut and be used and saved for later use as well? since there is only 4 of them and I was wondering if we could do that and if not, can we use a different adhesive strip, if this adhesive strip is only to protect the well from residues or the contamination issue?”

A:
Yes the adhesive strip can be cut and saved for future use.
Pls seal it well when using it. Pls let me know if you have any further questions. Thank you.
Q:

Please provide the following information regarding Rat Tyrosine Hydroxylase, TH ELISA Kit (CSB-E13102r)
a. Is there a predefined protocol on the SpectraMax M3 plate reader for this assay for wavelength correction?
b. Customer would like to know if the lab recommends 540 or 570 nm for the wavelength correction?
c. Also, the plate contains 12 strips of 8 wells, correct?
d. Also, can you please clarify if the kit is good for only one month at 2-8 degC? Or can it be stored longer?

A:
Thanks for your inquiry.
CSB-E13102r
The detection wavelength should be 450nm, and the correction wavelength can be 540 or 570nm.
We suggest you set dual wavelength( detection wavelength and correction wavelenth) to test the samples.
This kit is 8wells× 12 strips;
About storage conditions pls kindly refer to "STORAGE" in the manual, which is sent to you by e-mail.
Q:

Thank you for your response.I also would like to know if the lab recommends path length check on the plate reader?

A:
Thanks for your further message. The path length check on the plate reader is not needed.
Pls note that the detection wavelength must be 450nm, and the correction wavelength can be 540 or 570 or 630nm.Pls let me know if you have any further questions. Thank you.

Target Background

Function
(From Uniprot)
Plays an important role in the physiology of adrenergic neurons. Positively regulates the regression of retinal hyaloid vessels during postnatal development.
Gene References into Functions
  1. In hepatic encephalopathy model rats, a reduction in TH-positive neurons was seen at both 1 and 6 months in the substantia nigra pars compacta (SNc), as well as a reduction in TH-positive fibres in the striatum. This was coincident with the appearance of alpha-synuclein-immunoreactive neurons in the SNc; some of the TH-positive neurons also showed alpha-synuclein immunoreactivity. PMID: 28984618
  2. Study demonstrates that the local synthesis of Tyrosine Hydroxylase can modulate the levels of catecholamines in the axon and nerve terminal, and raises the possibility that trafficking and local synthesis of Tyrosine Hydroxylase mRNA may act to facilitate the restoration of neurotransmitter levels under conditions of prolonged release. PMID: 28630892
  3. Objective of the present study was to determine whether the lack of activity-stimulating effects of amphetamine in hyperactive rats is associated with changes in tyrosine hydroxylase (TH) activity. Study confirmed that changes in TH activity may be causally involved in the loss of amphetamine's locomotor-stimulating effects under conditions of impaired DAT activity. PMID: 28750831
  4. Study compared showing a clear-cut profile of either gambling proneness or aversion, found a selective down-regulation of i) serotonin transporter in prefrontal cortex, ii) tyrosine hydroxylase in ventral striatum, iii) dopamine transporter in lymphocytes was evidenced in "gambler" vs "non-gambler" rats. PMID: 27789384
  5. Our results suggest that HMGB1 upregulates TH expression to maintain dopaminergic neuronal function via activating RAGE, which is dependent on JNK phosphorylation. PMID: 28887039
  6. Data suggest that TH phosphorylated at Ser-31 co-distributes with Golgi complexes and synaptic-like vesicles in rat and human dopaminergic neurons/cell lines; Ser-31 phosphorylation may regulate TH subcellular localization by enabling its transport along microtubules, notably toward the projection terminals. PMID: 28637871
  7. tyrosine hydroxylase and dopamine beta-hydroxylase-was evaluated in the organ of Zuckerkandl of rats in the critical period of morphogenesis PMID: 28726094
  8. results provide the first direct evidence that TH mRNA is trafficked to the axon and that the mRNA is locally translated. These findings raise the interesting possibility that the biosynthesis of the catecholamine neurotransmitters is locally regulated in the axon and/or presynaptic nerve terminal PMID: 27095027
  9. these data demonstrate a novel interaction between Hsc70 and TH that regulates the activity and localization of the enzyme to synaptic vesicles, suggesting an important role for Hsc70 in dopamine homeostasis. PMID: 27365397
  10. Exercise-Mediated Increase in Nigral Tyrosine Hydroxylase in aging rats. PMID: 26599339
  11. LPS significantly increased the supernatant TNF-alpha and IL-6 concentrations and reduced TH protein expression in PC12 cells, while silencing Nogo-A could block these effects PMID: 26583134
  12. that deep brain stimulation of the anterior nucleus of the thalamus induced substantial higher numbers of TH-immunoreactive cells in the VTA PMID: 25074751
  13. Activation of proteasome degradation by IU-1, which is an inhibitor for the deubiquitinating activity of USP14, decreased only the quantity of TH molecules phosphorylated at their Ser19. PMID: 26969276
  14. Data show that Fischer 344 rats displayed greater tyrosine hydroxylase and dopamine transporter levels in the frontal cortex compared to the Lewis rats. PMID: 26658810
  15. Trx-1 is involved in TH expression induced by epinephrine in PC12 cells PMID: 25957836
  16. Dopamine deficiency induces compensatory activation of tyrosine hydroxylase via phosphorylation at 40Ser through D2-autoreceptor and PKA-mediated pathways. PMID: 26225746
  17. Data suggest that, in the Fe(II) catalytic center of TyrH, one water molecule coordinates with tyrosine, nitric oxide, and 6-methyltetrahydropterin. PMID: 26024204
  18. Atrial natriuretic peptide mediates negative regulation of TH gene expression via stimulation of PDE2-dependent cAMP breakdown in PC12 cells. PMID: 24837549
  19. These results demonstrate asymmetry in noradrenegic genes (Th, NET, and VMAT2) in the right and left stellate ganglia during stress and provide molecular evidence to help explain the difference in response to the stress. PMID: 24480406
  20. there exists possible interrelation between TH and tryptophan hydroxylase gene expression and epigenetic histone acetylation in chronic unpredictable stress-induced depressive rats, which at least partly contributes to the etiology of depression. PMID: 24495952
  21. acute footshock stress leads to activation of TH in the locus coeruleus, pre-synaptic terminals in the prefrontal cortex and adrenal medullary chromaffin PMID: 24117713
  22. Activation of Beclin 1-dependent autophagy may inhibit the loss of TH-positive neurons. PMID: 23868341
  23. Disruption of the blood-brain barrier results in increased activation of Th-containing cells by circulating angiotensin II. PMID: 23608099
  24. No changes were observed in gene expression and protein levels of TH in the right hippocampus in chronic stress, whereas expression of catecholamine synthesizing enzyme was elevated in the left hippocampus. PMID: 24317346
  25. expression of soluble GFRalpha1 in substantia nigra may be a key component in locomotor activity regulation through its influence over TH regulation and dopamine biosynthesis. PMID: 23321789
  26. The pattern of aging differs between male and female rats both on performance of a delayed alternation task and in the density of TH fibers in the medial prefrontal cortex. PMID: 23327742
  27. These results show that phosphorylation of TH at Ser31 and Ser40 is increased in carotid body glomus cells by short-term hypoxia PMID: 23153692
  28. Saturation mutagenesis of Asp425 results in large (up to 10(4)) decreases in the V(max) and V(max)/K(tyr) values for tyrosine hydroxylation, but only small decreases or even increases in the V(max) and V(max)/K(phe) values for phenylalanine hydroxylation. PMID: 23368961
  29. These findings imply that tyrosine hydroxylase activation of the ERK1/2 signal pathway could play an important role in ouabain-induced hyperactivity of rats, a mania model. PMID: 21871514
  30. albumin prevents 6-hydroxydopamine-induced loss of tyrosine hydroxylase PMID: 22815976
  31. Glucocorticoids are capable of inducing tyrosine hydroxylase at both transcriptional and enzyme activity levels in the brainstem of near-term fetuses. PMID: 22212880
  32. CNTF decreased TH enzyme levels, thereby increasing the specific activity of the enzyme PMID: 22372951
  33. Substance P, TH and serotonin are not always co-localized in terminals forming inputs with laryngeal motoneurons; they arise from separate subpopulations of neurons. PMID: 21689789
  34. Early life stress has enduring effects on the populations of midbrain TH-expressing dopaminergic neurons (especially in female rats), which are critically important for dopamine-regulated brain function throughout ontogenesis. PMID: 21396433
  35. TH regulation in the midbrain may be critical for dopamine bioavailability to influence behavior. PMID: 22242182
  36. Results compare the effect of a brief interruption then resumption of suckling on the temporal program of tyrosine hydroxylase and enkephalin regulation in lactating rats. PMID: 21125428
  37. Dopamine metabolite 3,4-dihydroxyphenylacetaldehyde covalently modifies and inhibits tyrosine hydroxylase activity. PMID: 21514317
  38. The decrease in TH content in the substantia nigra compacta correlated with the aging of rats. PMID: 21818658
  39. Stress did not alter tyrosine hydroxylase expression in heart ventricles. PMID: 21777029
  40. Measuring TH phosphorylation at Ser19, Ser31, and Ser40 over time can therefore be used as a sensitive index to differentiate the effects of different stressors on catecholaminergic cells. PMID: 21767616
  41. Endothelin 1 is involved in the long-term regulation of tyrosine hydroxylase, leading to an increase in catecholaminergic activity implicated in diverse pathologies involving the olfactory bulbs. PMID: 21129429
  42. alpha2-Adrenoceptors do not play a substantial role in regulating the tyrosine hydroxylase phenotype of hypothalamic magnocellular neurons during osmotic challenge induced by vasopressin deficiency. PMID: 20814066
  43. Data show that after stress, the expression of tyrosine hydroxylase was decreased in amygdale, and the levels of glucocorticoid and sphingosine 1-phosphate were elevated in serum. PMID: 21076868
  44. These findings suggest that the regulation of tyrosine hydroxylase phosphorylation in different catecholamine-producing cells varies considerably and is dependent on both the nature of the stressor and the time at which the response is analysed. PMID: 20811774
  45. This study suggested that the expression of either TH mRNA or TH protein was subject to the mediation of different transmembrane receptor combinations of GDNF. PMID: 20884338
  46. The expressions of TH mRNA and the immunoreactivity for TH were not altered in the CB of rats exposed to hypercapnia. It is suggested that CO(2) does not affect TH expression in the CB, and that it inhibits hypoxia-enhanced TH expression. PMID: 20620242
  47. transcriptional mechanisms are involved in mediating the immobilization stress triggered elevation in TH gene expression in the superior cervical ganglia. PMID: 20102321
  48. results indicate that, under hypoxic conditions, TH (a key factor in systemic adaptation to reduced oxygen availability) is not regulated by HIF-1, the primary modulator of the response to hypoxia, but by the A2aR-mediated signalling pathway PMID: 20143408
  49. The renal protective effect of ginsenosides against acute renal failure was accompanied by up-regulation of tyrosine hydroxylase in the locus coeruleus. PMID: 19249909
  50. Data show that inactivation of tyrosine hydroxylase in hypothalamic neuroendocrine dopamine neurons is required for suckling-induced prolactin and ACTH responses. PMID: 20170714

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Subcellular Location
Cytoplasm, perinuclear region.
Protein Families
Biopterin-dependent aromatic amino acid hydroxylase family
Database Links
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