Invitrogen™ Amplex™ Red Neuraminidase (Sialidase) Assay Kit

Catalog No.	Quantity
A22178	400 Assays

Catalog No. A22178 Supplier Invitrogen™ Supplier No. A22178

Description

Format allows for multiple time point measurements

The Amplex Red Neuraminidase (Sialidase) Assay Kit provides a sensitive and simple method for detecting neuraminidase activity in serum and in purified enzyme systems using either a fluorescence microplate reader or fluorometer. Neuraminidase (also known as sialidase) is a very common enzyme that hydrolyzes terminal sialic acid residues on polysaccharide chains, most often exposing a galactose residue. Neuraminidase plays a key role in the invasion of target cells and the subsequent replication of the influenza virus, making neuraminidase an important target for influenza drug development. The Amplex Red Neuraminidase (Sialidase) Assay Kit provides an ultrasensitive method for detecting neuramimidase activity. This assay utilizes Amplex Red reagent to detect hydrogen peroxide generated by galactose oxidase oxidation of desialiated galactose, the end result of neuraminidase action. Then the hydrogen peroxide, in the presence of horseradish peroxidase (HRP), reacts in a 1:1 stoichiometric ratio with Amplex Red reagent to generate the red-fluorescent oxidation product resorufin. Because resorufin has absorption and fluorescence emission maxima of approximately 571nm and 585nm, respectively, there is little interference from autofluorescence in most biological samples.

• Detect neuraminidase levels as low as 0.2mU/mL
• Format allows for multiple time point measurements
• Designed for minimal autofluorescence interference
• Detect neuraminidase activity in biological samples such as serum
• Custom assay design and packaging are also available

Use Amplex Red Assays for a Broad Range of Investigations

A wide variety of validated Amplex Red assays are available for studying cell signaling and lipids, neurobiology, inflammation and immune function, and metabolism. We also offer Amplex UltraRed Reagent, a second-generation reagent providing greater sensitivity and brighter fluorescence, and the Amplex Red/UltraRed Stop Reagent. The Amplex Red/UltraRed Stop Reagent provides convenience and control by allowing the fluorescence signal-generating reaction to be terminated at a user-determined time point. After addition of the stop reagent, the fluorescence signal remains stable for at least three hours.

Enzyme and Protein Activity Assays, Glycosidase, Neuraminidase, and Related Enzyme Activity, Proteins, Expression, Isolation and Analysis

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
• For Use With (Application): Neuraminidase (Sialidase) Assay
• For Use With (Equipment): Microplate Reader, Spectrophotometer, Fluorometer
• Product Type: Amplex Red Assay Kit
• Dye Type: Other Label(s) or Dye(s)
• Format: Tube(s), 96-well plate
• Product Line: Amplex
• Quantity: 400 Assays
• Shipping Condition: Room Temperature

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.

How does the Amplex Red Neuraminidase (Sialidase) Assay kit detect neuramimidase activity?

The assay utilizes Amplex Red to detect H₂O₂ generated by galactose oxidase oxidation of desialiated galactose, the result of neuraminidase activity. In the presence of HRP, H₂O₂ reacts with a 1:1 stiochiometry with Amplex Red reagent to generate the red-fluorescent oxidation product, resorufin.

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