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alanyl-tRNA synthetase Cell Lysate | AARS cell lysate

Human alanyl-tRNA synthetase Baculovirus-Insect Overexpression Lysate

Gene Names
AARS; CMT2N
Applications
Western Blot
Synonyms
alanyl-tRNA synthetase; Human alanyl-tRNA synthetase Baculovirus-Insect Overexpression Lysate; Human AARS/alanyl-tRNA synthetase Insect Cell Lysate (WB positive control); Human CMT2N Overexpression Lysate; AARS 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
987aa
Sequence
Met1-Asn968
Applicable Applications for AARS 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.
3. Store the lyophilized cell lysate at 4 degree C. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80 degree C.
Sequence Construction
A DNA sequence encoding the human AARS (NP_001596.2) (Met1-Asn968) was expressed with a polyhistidine tag at the N-terminus.
Preparation Method
Cell lysate was prepared by homogenization 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 AARS cell lysate
Alanyl-tRNA synthetase (AARS) belongs to the family of ligases, specifically those forming carbon-oxygen bonds in aminoacyl-tRNA and related compounds. This enzyme participates in alanine and aspartate metabolism and aminoacyl-tRNA biosynthesis. Alanyl-tRNA synthetase (AlaRS) catalyzes synthesis of Ala-tRNA (Ala) and hydrolysis of mis-acylated Ser-and Gly-tRNA (Ala) at 2 different catalytic sites. Their role is not confined to catalyze the attachment of amino acids to transfer RNAs and thereby establish the rules of genetic code by virtue of matching the nucleotide triplet of anticodon with cognate amino acid. Under apoptotic conditions in cell culture, the full-length enzyme is secreted, and the two cytokine activities can be generated by leukocyte elastase, an extracellular protease. Secretion of this tRNA synthetase may contribute to apoptosis both by arresting translation and producing needed cytokines. This protein could be an attractive target of drugs against bacterial, fungal and parasitic infections.

This Human alanyl-tRNA synthetase overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of alanyl-tRNA synthetase protein from the overexpression lysate was verified.
References
Wakasugi K, et al. (1999) Two Distinct Cytokines Released from a Human Aminoacyl-tRNA Synthetase. Science. 284 (5411): 147-51.Sokabe M, et al. (2009) The structure of alanyl-tRNA synthetase with editing domain. Proc Natl Acad Sci. 106 (27): 11028-33.Skupinska M, et al. (2009) AARS--the etiological factor and the attractive target of many disorders. Postepy Biochem. 55 (4): 373-84.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
16
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
109,317 Da
NCBI Official Full Name
alanine--tRNA ligase, cytoplasmic
NCBI Official Synonym Full Names
alanyl-tRNA synthetase
NCBI Official Symbol
AARS
NCBI Official Synonym Symbols
CMT2N
NCBI Protein Information
alanine--tRNA ligase, cytoplasmic; AARS; alaRS; alanine tRNA ligase 1, cytoplasmic; alanyl-tRNA synthetase, cytoplasmic; renal carcinoma antigen NY-REN-42
UniProt Protein Name
Alanine--tRNA ligase, cytoplasmic
UniProt Gene Name
AARS
UniProt Entry Name
SYAC_HUMAN

NCBI Description

The human alanyl-tRNA synthetase (AARS) belongs to a family of tRNA synthases, of the class II enzymes. Class II tRNA synthases evolved early in evolution and are highly conserved. This is reflected by the fact that 498 of the 968-residue polypeptide human AARS shares 41% identity witht the E.coli protein. tRNA synthases are the enzymes that interpret the RNA code and attach specific aminoacids to the tRNAs that contain the cognate trinucleotide anticodons. They consist of a catalytic domain which interacts with the amino acid acceptor-T psi C helix of the tRNA, and a second domain which interacts with the rest of the tRNA structure. [provided by RefSeq, Jul 2008]

Uniprot Description

AARS: Catalyzes the attachment of alanine to tRNA(Ala) in a two-step reaction: alanine is first activated by ATP to form Ala- AMP and then transferred to the acceptor end of tRNA(Ala). Also edits incorrectly charged tRNA(Ala) via its editing domain. Defects in AARS are the cause of Charcot-Marie-Tooth disease type 2N (CMT2N). It is an axonal form of Charcot-Marie-Tooth disease, a disorder of the peripheral nervous system, characterized by progressive weakness and atrophy, initially of the peroneal muscles and later of the distal muscles of the arms. Charcot-Marie-Tooth disease is classified in two main groups on the basis of electrophysiologic properties and histopathology: primary peripheral demyelinating neuropathies(designated CMT1 when they are dominantly inherited) and primary peripheral axonal neuropathies (CMT2). Neuropathies of the CMT2 group are characterized by signs of axonal regeneration in the absence of obvious myelin alterations, normal or slightly reduced nerve conduction velocities, and progressive distal muscle weakness and atrophy. Nerve conduction velocities are normal or slightly reduced. Belongs to the class-II aminoacyl-tRNA synthetase family.

Protein type: RNA-binding; Ligase; EC 6.1.1.7; Translation

Chromosomal Location of Human Ortholog: 16q22

Cellular Component: membrane; cytoplasm; cytosol

Molecular Function: amino acid binding; zinc ion binding; ATP binding; alanine-tRNA ligase activity; tRNA binding

Biological Process: protein folding; unfolded protein response; tRNA processing; regulation of translational fidelity; cerebellar Purkinje cell layer development; tRNA modification; response to amino acid stimulus; tRNA aminoacylation for protein translation; hair follicle development; gene expression; negative regulation of neuron apoptosis; neuromuscular process controlling balance; alanyl-tRNA aminoacylation

Disease: Epileptic Encephalopathy, Early Infantile, 29; Charcot-marie-tooth Disease, Axonal, Type 2n

Research Articles on AARS

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Product Notes

The AARS aars (Catalog #AAA8114254) 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 987aa. AAA Biotech's alanyl-tRNA synthetase 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 AARS aars 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: Met1-Asn96 8. It is sometimes possible for the material contained within the vial of "alanyl-tRNA synthetase, 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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