Invitrogen™ Amplex™ Acetylcholine/Acetlycholinesterase Assay Kit

Catalog No.	Quantity
A12217	500 Assays

Catalog No. A12217 Supplier Invitrogen™ Supplier No. A12217

Description

Detect activity levels as low as 0.002U/mL in one hour

The Amplex Red Acetylcholine/Acetylcholinesterase Assay Kit provides an ultrasensitive method for detecting acetylcholinesterase (AChE) activity in a fluorescence microplate reader or fluorometer. AChE activity is monitored indirectly using 10-acetyl-3,7-dihydroxyphenoxazine (Amplex Red reagent), a sensitive fluorogenic probe for hydrogen peroxide. AChE converts the acetylcholine substrate to choline, which is then oxidized by choline oxidase to betaine and hydrogen peroxide. In the presence of horseradish peroxidase, hydrogen peroxide reacts with the Amplex Red reagent in a 1:1 stoichiometric ratio to generate the highly fluorescent product resorufin.

• Little interference from autofluorescence in most biological samples
• Detect AChE activity levels as low as 0.002U/mL in one hour
• Detect acetylcholine levels as low as 0.3μM using excess AChE
• Achieve detection ranges of 0.3μM to 100μM acetylcholine
• Format allows for multiple time point measurements
• Designed for minimal autofluorescence interference
• Custom assay design and packaging are also available

Drug Discovery and Development, Enzyme and Protein Activity Assays, Proteins, Expression, Isolation and Analysis, Target and Lead Identification and Validation

Order Info

Shipping Condition: Room temperature

Specifications

• Content And Storage: Store in freezer -5°C to -30°C and protect from light.
• Detection Method: Fluorescence Intensity
• For Use With (Application): Acetylcholine/Acetlycholinesterase Assay
• For Use With (Equipment): Fluorescence Microplate Reader
• Product Type: Amplex Red Assay Kit
• Product Line: Amplex
• Quantity: 500 Assays
• Shipping Condition: Room Temperature
• Substrate Properties: Molecular Substrate
• Substrate Type: HRP (Horseradish Peroxidase) Substrate, Acetylcholinesterase Substrate
• Target Enzyme: Acetylcholinesterase

Frequently Asked Questions (FAQs)

I'm using an Amplex Red kit, the reagent changes color to pink almost immediately in my own Krebs-Ringer buffer but not in HBSS. Why is this?

The components of Krebs-Ringer buffer (salts) should not cause oxidation of the Amplex reagent (which, in the presence of peroxidase and H₂O₂ oxidizes to resorufin, which is pink in color and fluorescent). Try water alone (the water used to make the Krebs-Ringer buffer). Since Hank's Buffered Saline Solution is typically purchased rather than made in the lab, it likely would not have the same contaminant. Another option is to degas the buffer prior to use to removed dissolved oxygen radicals.

Can Amplex Red Assays be performed using cell lysates?

This is not recommended. The presence of endogenous proteases can complicate the assay by degrading the horseradish peroxidase (HRP). Endogenous peroxidases and antioxidants can modify the H₂O₂ required for the reaction, competing with HRP (and catalase) for the substrate.

The Amplex Red Assays are best performed with either purified enzymes or extracted H₂O₂ in a defined buffer system, extracellular solutions or body fluids (media, serum, etc.) that do not exhibit high levels of endogenous protease or oxidase activity and do not contain antioxidants.

For Research Use Only. Not for use in diagnostic procedures.