Acetylcholinesterase/ACHE Protein, Mouse (HEK293, His)

Acetylcholinesterase (ACHE) proteins exhibit multiple functions, including acetylcholinesterase activity and protein self-association. It regulates acetylcholine processing, receptor signaling and recycling. Acetylcholinesterase/ACHE Protein, Mouse (HEK293, His) is the recombinant mouse-derived Acetylcholinesterase/ACHE protein, expressed by HEK293 , with C-10*His labeled tag.

Acetylcholinesterase (ACHE) protein, with its diverse functional repertoire, exhibits acetylcholinesterase activity, identical protein binding activity, and the ability for protein self-association. Its involvement in positive regulation of cold-induced thermogenesis positions it upstream of critical processes such as acetylcholine catabolic processes, acetylcholine receptor signaling pathways, and the regulation of receptor recycling. ACHE is found in various cellular components, including the basement membrane, cell surface, and neuromuscular junction. Its expression spans across multiple structures, including the alimentary system, genitourinary system, musculature, nervous system, and sensory organs. Notably, human orthologs of ACHE are implicated in significant health conditions such as Alzheimer's disease, epilepsy, and myasthenia gravis, highlighting the importance of this protein in various physiological contexts. Broad expression patterns further emphasize its fundamental role in multiple tissues, particularly in the cerebellum and thymus.

Measured by its ability to cleave Acetylthiocholine. The specific activity is > 250 nmoL/min/μg, as measured under the described conditions.

Materials
Assay buffer: 0.1 M sodium phosphate with 0.05% Brij-35, pH 7.5
Acetylcholinesterase/ACHE Protein, Mouse (HEK293, His) (HY-P72811)
DTNB (HY-15915): Dissolve in DMSO to make a 10 mM solution, then dilute with buffer to 100 μM
Substrate mixture: First dilute ATC in DMSO to 20 mM, then dilute ATC to 200 μM with 100 μM DTNB to prepare the substrate mixture

Assay Procedure
1. Dilute Mouse ACHE protein to 0.02 μg/mL with buffer;
2. Add 50 μL of 0.02 μg/mL Mouse ACHE protein buffer to a 96-well plate, then add 50 μL of substrate to start the reaction. Include three replicates per group and a blank control containing 50 μL of buffer and 50 μL of substrate mixture.
3. Zero the microplate reader using the OD value of the blank group, and read the absorbance at 405 nm in kinetic mode for 5 minutes.
4. Calculate the specific activity:

Mouse

HEK293

C-10*His

P21836-1/NP_033729.1 (E32-L614)

11423  [NCBI]

Full Length of Isoform-1 Mature Protein

EGREDPQLLVRVRGGQLRGIRLKAPGGPVSAFLGIPFAEPPVGSRRFMPPEPKRPWSGVLDATTFQNVCYQYVDTLYPGFEGTEMWNPNRELSEDCLYLNVWTPYPRPASPTPVLIWIYGGGFYSGAASLDVYDGRFLAQVEGAVLVSMNYRVGTFGFLALPGSREAPGNVGLLDQRLALQWVQENIAAFGGDPMSVTLFGESAGAASVGMHILSLPSRSLFHRAVLQSGTPNGPWATVSAGEARRRATLLARLVGCPPGGAGGNDTELIACLRTRPAQDLVDHEWHVLPQESIFRFSFVPVVDGDFLSDTPEALINTGDFQDLQVLVGVVKDEGSYFLVYGVPGFSKDNESLISRAQFLAGVRIGVPQASDLAAEAVVLHYTDWLHPEDPTHLRDAMSAVVGDHNVVCPVAQLAGRLAAQGARVYAYIFEHRASTLTWPLWMGVPHGYEIEFIFGLPLDPSLNYTTEERIFAQRLMKYWTNFARTGDPNDPRDSKSPQWPPYTTAAQQYVSLNLKPLEVRRGLRAQTCAFWNRFLPKLLSATDTLDEAERQWKAEFHRWSSYMVHWKNQFDHYSKQERCSDL

Approximately 80 kDa

• 
  ≥ 95%, as determined by reducing SDS-PAGE.

Lyophilized powder.

Lyophilized from a 0.2 μm filtered solution of PBS, pH 7.4. Normally 5 % - 8 % trehalose, mannitol and 0.01% Tween 80 are added as protectants before lyophilization.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O.

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.