Acetylcholinesterase/ACHE Protein, Human (CHO, His)

Acetylcholinesterase (ACHE) protein is pivotal in neurotransmission, rapidly hydrolyzing acetylcholine in the synaptic cleft to terminate signal transduction at the neuromuscular junction. Beyond neurotransmitter regulation, ACHE is implicated in neuronal apoptosis, indicating a broader role in cellular processes associated with programmed cell death. Acetylcholinesterase/ACHE Protein, Human (CHO, His) is the recombinant human-derived Acetylcholinesterase/ACHE protein, expressed by CHO , with C-6*His labeled tag.

Acetylcholinesterase (ACHE) protein plays a crucial role in neurotransmission by rapidly hydrolyzing the acetylcholine neurotransmitter in the synaptic cleft, thereby facilitating the termination of signal transduction at the neuromuscular junction. Beyond its essential function in neurotransmitter regulation, ACHE is implicated in neuronal apoptosis, suggesting a broader role in cellular processes related to programmed cell death.

Measured by its ability to cleave Acetylthiocholine that incubate at room temperature in kinetic mode for 5 minutes. The specific activity is ≥308.73 nmol/min/μg.

Materials
Acetylcholinesterase/ACHE Protein, Human (CHO, His) (HY-P79277)
Assay buffer: 0.1 M Sodium Phosphate with 0.05% Brij-35, pH 7.5
DTNB (HY-15915): Dissolve in DMSO to 10 mM, then dilute to 100 μM with assay buffer
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

Procedure
1. Dilute ACHE protein to 0.002 μg/mL and 0.02 μg/mL respectively with buffer.
2. Add 50 μL of ACHE protein buffer at different concentrations to a 96-well plate, then add 50 μL of substrate to initiate the reaction, with 3 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 min.
4. Calculate the specific activity:

Human

CHO

C-6*His

P22303-1 (E32-L614)

43  [NCBI]

Full Length of Isoform-1 Mature Protein

EGREDAELLVTVRGGRLRGIRLKTPGGPVSAFLGIPFAEPPMGPRRFLPPEPKQPWSGVVDATTFQSVCYQYVDTLYPGFEGTEMWNPNRELSEDCLYLNVWTPYPRPTSPTPVLVWIYGGGFYSGASSLDVYDGRFLVQAERTVLVSMNYRVGAFGFLALPGSREAPGNVGLLDQRLALQWVQENVAAFGGDPTSVTLFGESAGAASVGMHLLSPPSRGLFHRAVLQSGAPNGPWATVGMGEARRRATQLAHLVGCPPGGTGGNDTELVACLRTRPAQVLVNHEWHVLPQESVFRFSFVPVVDGDFLSDTPEALINAGDFHGLQVLVGVVKDEGSYFLVYGAPGFSKDNESLISRAEFLAGVRVGVPQVSDLAAEAVVLHYTDWLHPEDPARLREALSDVVGDHNVVCPVAQLAGRLAAQGARVYAYVFEHRASTLSWPLWMGVPHGYEIEFIFGIPLDPSRNYTAEEKIFAQRLMRYWANFARTGDPNEPRDPKAPQWPPYTAGAQQYVSLDLRPLEVRRGLRAQACAFWNRFLPKLLSATDTLDEAERQWKAEFHRWSSYMVHWKNQFDHYSKQDRCSDL

Approximately 65-80 kDa due to the glycosylation.

• 
  Greater than 95% as determined by reducing SDS-PAGE.

Lyophilized powder.

Lyophilized from a 0.22 μm filtered solution of 20 mM Tris, 150 mM NaCl, pH 7.4 or 20 mM Tris-HCl, 150 mM NaCl, pH 7.4, 8% trehalose.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

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.