The largest database of trusted experimental protocols

Amplite fluorimetric acetylcholinesterase assay kit

Manufactured by AAT Bioquest
Sourced in United States

The Amplite Fluorimetric Acetylcholinesterase Assay Kit is a laboratory product designed to measure the activity of the enzyme acetylcholinesterase. The kit provides a fluorescence-based method for quantifying acetylcholinesterase levels in various samples.

Automatically generated - may contain errors

4 protocols using amplite fluorimetric acetylcholinesterase assay kit

1

Fluorimetric Assay for Lung AChE Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols
For acetylcholinesterase (AChE) activity assay, mice lungs were rapidly dissected under deep anaesthetisation and frozen in liquid nitrogen. The frozen lung samples were homogenised in ice-cold phosphate buffer (50 mM, pH 7.4) using an Ultrasonic Homogenizer VP-050N (TAITEC, Tokyo, Japan). The lung homogenates were centrifuged at 11,000×g for 20 min, and the supernatants were aliquoted and stored at − 80 °C until assay. The concentration of total protein was determined using the Quick Start Bradford Protein Assay reagents (Bio-Rad, Tokyo, Japan) and bovine serum albumin standard (Bio-Rad). AChE activity was measured by duplicate analysis per sample using the Amplite Fluorimetric Acetylcholinesterase Assay Kit (AAT Bioquest, Sunnyvale, CA, USA) in accordance with the manufacturer’s instructions. The AChE activity of each sample was determined by the standard curve, and enzymatic activity was calculated as mU/mg protein.
+ Open protocol
+ Expand
2

Quantification of AChE Activity in PC12 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
The thiocholine produced from the hydrolysis of acetylthiocholine by the endogenous AchE enzyme in each sample was quantified by a fluorescence colorimetric assay to examine the effect of ILA on AchE activity, which is a biochemical marker for neuronal differentiation in PC12 cells [28 (link)]. In brief, PC12 cells were grown and treated for five consecutive days, as mentioned. The cells were then lysed with ice-cold NP-40 cell lysis buffer containing 150 mM NaCl, 50 mM Tris (pH 8.0), 2 mM EDTA (pH 8.0) and 1% (v/v) NP-40. AchE activity of the cell lysates was determined by the Amplite Fluorimetric Acetylcholinesterase Assay Kit (AAT Bioquest, Sunnyvale, CA, USA) according to the manufacturer’s instructions. Assay signals were read with a fluorescence absorbance microplate reader (SH-9000, Corona Electric, Ibaraki, Japan). at Ex/Em = 490/520 nm. The AchE activity was determined from the standard curve and normalized with the protein concentration in each sample. The protein concentration was determined by Pierce bicinchoninic acid (BCA) protein assay kit (Invitrogen, Paisley, UK) with bovine serum albumin as a standard.
+ Open protocol
+ Expand
3

Modulation of AChE Activity in PC12 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
PC12 cells (10,000 cells/mL per well) were cultured in collagen type IV-coated 24-well culture plates, as mentioned. The cells were then exposed to the antagonists of AhR receptor (ANF and CH223191) at a final concentration of 1 µM for 1 h prior to the treatment with NGF (25 ng/mL) and the test compounds (reference control, ILA or tryptophan at a final concentration of 100 nM) for five consecutive days. Control cells without ANF and CH223191 were also grown and treated under the same conditions. The medium and test compounds were replaced after three days. Following treatment, the cells were lysed with ice-cold NP-40 cell lysis buffer. AchE activity of the cell lysates was then determined by the Amplite Fluorimetric Acetylcholinesterase Assay Kit (AAT Bioquest, Sunnyvale, CA, USA) according to the manufacturer’s instructions. The AchE activity was determined as described above.
+ Open protocol
+ Expand
4

Acetylcholinesterase Activity in Zebrafish

Check if the same lab product or an alternative is used in the 5 most similar protocols
After co-treatment with AlCl3 and EUMF, zebrafish larvae at 6 dpf were killed by tricaine (Sigma-Aldrich, Darmstadt, Germany). Cold physiological saline was added to the larvae in a 2 mL tube at a ratio of 1:9 (mass:volume) without any additional water. Next, the samples were homogenized using automated tissue homogenization, followed by centrifuged at 2,500 rpm for 10 min at 0°C. The supernatant was collected for the assay. The enzyme activity of Ache was determined by using the Amplite™ Fluorimetric Acetylcholinesterase Assay Kit (AAT Bioquest, California, United States) according to the manufacturer’s instructions with a slight modification as follows. The acetylthiocholine reaction mixture was 50 μM.
The test samples addition added into the acetylthiocholine reaction mixture was also 50 μM. The fluorescence at Ex/Em = 490/520 was monitored.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!