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Tyrosine Hydroxylase Cell Lysate | TH cell lysate

Human Tyrosine Hydroxylase Baculovirus-Insect cells Overexpression Lysate

Gene Names
TH; TYH; DYT14; DYT5b
Applications
Western Blot
Synonyms
Tyrosine Hydroxylase; Human Tyrosine Hydroxylase Baculovirus-Insect cells Overexpression Lysate; Human TH/Tyrosine Hydroxylase Insect Cell Lysate (WB positive control); tyrosine hydroxylase; Human DYT14 Overexpression Lysate; Human DYT5b Overexpression Lysate; Human TYH Overexpression Lysate; TH cell lysate
Ordering
For Research Use Only!
Host
Baculovirus-Insect cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading buffer).
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Sequence Positions
515aa
Sequence
Pro2-Gly497
Applicable Applications for TH cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Human
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the human TH isoform 2 (P07101-3) (Pro 2-Gly 497) was fused with a polyhistidine tag at the N-terminus.
Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for TH cell lysate
Tyrosine hydroxylase (TH) is a rate-limiting enzyme in catecholamine synthesis. Tyrosine hydroxylase activity is modulated by protein-protein interactions with enzymes in the same pathway or the tetrahydrobiopterin pathway, structural proteins considered to be chaperones that mediate the neuron's oxidative state, and the protein that transfers dopamine into secretory vesicles. It is phosphorylated at serine (Ser) residues Ser8, Ser19, Ser31 and Ser40 in vitro. The phosphorylation of tyrosine hydroxylase at Ser19 or Ser8 has no direct effect on tyrosine hydroxylase activity. As tyrosine hydroxylase (TH) catalyses the formation of L-DOPA, the rate-limiting step in the biosynthesis of DA, the Parkinson's disease (PD) can be considered as a TH-deficiency syndrome of the striatum. A direct pathogenetic role of TH has also been suggested, as the enzyme is a source of reactive oxygen species (ROS) in vitro and a target for radical-mediated oxidative injury. Recently, it has been demonstrated that L-DOPA is effectively oxidized by mammalian Tyrosine hydroxylase in vitro, possibly contributing to the cytotoxic effects of DOPA.

This Human Tyrosine Hydroxylase overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of Tyrosine Hydroxylase protein from the overexpression lysate was verified.
References
Daubner SC, et al. (2011) Tyrosine hydroxylase and regulation of dopamine synthesis. Arch Biochem Biophys. 508(1): 1-12.Dunkley PR, et al. (2004) Tyrosine hydroxylase phosphorylation: regulation and consequences. J Neurochem. 91(5): 1025-43.Haavik J, et al. (1998) Tyrosine hydroxylase and Parkinson's disease. Mol Neurobiol. 16(3): 285-309.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
58,600 Da
NCBI Official Full Name
tyrosine 3-monooxygenase isoform b
NCBI Official Synonym Full Names
tyrosine hydroxylase
NCBI Official Symbol
TH
NCBI Official Synonym Symbols
TYH; DYT14; DYT5b
NCBI Protein Information
tyrosine 3-monooxygenase; dystonia 14; tyrosine 3-hydroxylase
UniProt Protein Name
Tyrosine 3-monooxygenase
UniProt Gene Name
TH
UniProt Synonym Gene Names
TYH; TH
UniProt Entry Name
TY3H_HUMAN

NCBI Description

The protein encoded by this gene is involved in the conversion of tyrosine to dopamine. It is the rate-limiting enzyme in the synthesis of catecholamines, hence plays a key role in the physiology of adrenergic neurons. Mutations in this gene have been associated with autosomal recessive Segawa syndrome. Alternatively spliced transcript variants encoding different isoforms have been noted for this gene. [provided by RefSeq, Jul 2008]

Uniprot Description

TH: an enzyme involved in the conversion of phenylalanine to dopamine. As the rate-limiting enzyme in the synthesis of catecholamines, tyrosine hydroxylase has a key role in the physiology of adrenergic neurons. Four splice variant isoforms have been described.

Protein type: Oxidoreductase; Endoplasmic reticulum; Mitochondrial; Vesicle; Amino Acid Metabolism - tyrosine; EC 1.14.16.2

Chromosomal Location of Human Ortholog: 11p15.5

Cellular Component: internal side of plasma membrane; neuron projection; smooth endoplasmic reticulum; mitochondrion; melanosome membrane; dendrite; terminal button; perikaryon; cytosol; synaptic vesicle; cytoplasm; cytoplasmic vesicle; nucleus

Molecular Function: protein domain specific binding; amino acid binding; protein binding; enzyme binding; ferric iron binding; ferrous iron binding; dopamine binding; oxygen binding; tyrosine 3-monooxygenase activity

Biological Process: heart morphogenesis; heart development; response to lipopolysaccharide; dopamine biosynthetic process; phytoalexin metabolic process; norepinephrine biosynthetic process; dopamine biosynthetic process from tyrosine; catecholamine biosynthetic process; response to electrical stimulus; epinephrine biosynthetic process; response to pyrethroid; neurotransmitter biosynthetic process; response to corticosterone stimulus; response to light stimulus; anatomical structure morphogenesis; phthalate metabolic process; mating behavior; social behavior; eye photoreceptor cell development; organ morphogenesis; response to ethanol; circadian sleep/wake cycle; response to zinc ion; cerebral cortex development; response to activity; response to water deprivation; synaptic transmission, dopaminergic; response to peptide hormone stimulus; locomotory behavior; fatty acid metabolic process; response to salt stress; response to estradiol stimulus; regulation of heart contraction; sensory perception of sound; visual perception; glycoside metabolic process; response to nutrient levels; terpene metabolic process; sphingolipid metabolic process; eating behavior; response to amphetamine; multicellular organismal aging; isoquinoline alkaloid metabolic process; response to herbicide; learning; response to ether; synaptic vesicle amine transport; memory; pigmentation; response to hypoxia; embryonic camera-type eye morphogenesis

Disease: Segawa Syndrome, Autosomal Recessive

Research Articles on TH

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Product Notes

The TH th (Catalog #AAA8114029) is a Cell Lysate produced from Baculovirus-Insect cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 515aa. AAA Biotech's Tyrosine Hydroxylase can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the TH th for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Pro2-Gly49 7. It is sometimes possible for the material contained within the vial of "Tyrosine Hydroxylase, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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