AAT Bioquest

Amplite® Fluorimetric Neuraminidase Assay Kit *Blue Fluorescence*

Neuraminidase dose response was measured in a 96-well black plate with Amplite® Fluorimetric Neuraminidase Assay Kit using a Gemini fluorescence microplate reader (Molecular Devices).
Neuraminidase dose response was measured in a 96-well black plate with Amplite® Fluorimetric Neuraminidase Assay Kit using a Gemini fluorescence microplate reader (Molecular Devices).
Neuraminidase dose response was measured in a 96-well black plate with Amplite® Fluorimetric Neuraminidase Assay Kit using a Gemini fluorescence microplate reader (Molecular Devices).
Ordering information
Catalog Number
Unit Size
Add to cart
Additional ordering information
InternationalSee distributors
Bulk requestInquire
Custom sizeInquire
ShippingStandard overnight for United States, inquire for international
Request quotation
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22


Neuraminidases, also called sialidases, are glycoside hydrolase enzymes that catalyze the hydrolysis of terminal sialic acid residues and neuraminic acid. The most commonly known neuraminidase is the viral neuraminidase. The cleavage of linkage between sialic acid and adjacent sugar residue permits the transport of the virus through mucin and destroys the haemagglutinin receptor on the host cell, thus allowing elution of progeny virus particles from infected cells. Neuraminidase promotes influenza virus release from infected cells and facilitates virus spread within the respiratory tract. Thus, it is an important target for influenza drug development. The detection of neuraminidase and screening its inhibitors is one of the essential tasks for investigating biological processes and prevention of influenza infection. There are a few assay kits available for detecting neuraminidase, but all the commercial available kits are tedious to use. Our Amplite® Fluorimetric Neuraminidase Assay Kit provides a sensitiveand robust fluorimetric assay to detect neuraminidase that exists either in cells or biological samples. The non-fluorescent neuraminidase substrate becomes strongly fluorescent upon neuraminidase cleavage. The kit can detect as little as 0.3 mU/mL neuraminidase in a 100 µL assay volume. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step. The signal can be easily read by a fluorescence microplate reader.


Fluorescence microplate reader

Excitation320 nm
Emission460 nm
Cutoff420 nm
Recommended plateSolid black


Example protocol


Protocol summary

  1. Prepare Neuraminidase working solution (50 µL)
  2. Add Neuraminidase standards or test samples (50 µL)
  3. Incubate at 37°C or room temperature for 1 - 2 hours
  4. Monitor fluorescence increase at Ex/Em = 320/460 nm (Cutoff = 420 nm)

Important notes
Thaw all the kit components at room temperature before starting the experiment.


Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

1. Neuraminidase standard solution (2U/mL):
Add 50 µL of ddH2O into the vial of Neuraminidase Standard (Component C) to make approximately 2 U/mL Neuraminidase standard solution.

2. FluLite™ Blue stock solution (200X):
Add 50 µL of ddH2O into the vial of FluLite™ Blue (Component A) to make 200X stock solution.


Neuraminidase standard

For convenience, use the Serial Dilution Planner: https://www.aatbio.com/tools/serial-dilution/12602

Add 10 µL of 2 U/mL Neuraminidase standard stock solution to 990 µL of Assay Buffer (Component B) to generate 20 mU/mL Neuraminidase standard (NA7). Take 20 mU/mL Neuraminidase standard solution and perform 1:2 serial dilutions to get serially diluted Neuraminidase standards (NA6 - NA1) with Assay Buffer (Component B). Note: Diluted Neuraminidase standard solution is unstable. Use within 4 hours.


Add 25 μL of 200X FluLite™ Blue stock solution into 5 mL of Assay Buffer (Component B) and mix well to prepare Neuraminidase working solution.


Table 1. Layout of neuraminidase standards and test samples in a solid black 96-well microplate. NA= Neuraminidase Standards (NA1 - NA7, 0.312 to 20 mU/mL), BL=Blank Control, TS=Test Samples.


Table 2. Reagent composition for each well.

NA1 - NA750 µLSerial Dilution (0.312 to 20 mU/mL)
BL50 µLBlank Control
TS50 µLtest sample
  1. Prepare Neuraminidase standards (NA), blank controls (BL), and test samples (TS) according to the layout provided in Tables 1 and 2. For a 384-well plate, use 25 µL of reagent per well instead of 50 µL.

  2. Add 50 µL of Neuraminidase working solution to each well of Neuraminidase standard, blank control, and test samples to make the total Neuraminidase assay volume of 100 µL/well. For a 384-well plate, add 25 µL of Neuraminidase working solution into each well instead, for a total volume of 50 µL/well.

  3. Incubate the reaction at 37°C or room temperature for 1 to 2 hours, protected from light. Note: 37°C incubation gives better results.

  4. Monitor the fluorescence increase at Ex/Em = 320/460 nm (Cutoff = 420 nm) with a fluorescence microplate reader.


Open in Advanced Spectrum Viewer

Spectral properties



View all 50 references: Citation Explorer
Rapid screening, identification, and purification of neuraminidase inhibitors from Lithospermum erythrorhizon Sieb.et Zucc. by ultrafiltration with HPLC-ESI-TOF-MS combined with semipreparative HPLC
Authors: Zhang M, Zhao H, Zhao Z, Yan H, Lv R, Cui L, Yuan J, Wang D, Geng Y, Liu D, Wang X.
Journal: J Sep Sci (2016): 2097
Neuraminidase inhibiting antibody responses in pigs differ between influenza A virus N2 lineages and by vaccine type
Authors: S, undefined and bulte MR, Gauger PC, Kitikoon P, Chen H, Perez DR, Roth JA, Vincent AL.
Journal: Vaccine (2016): 3773
Development of a surface plasmon resonance assay to measure the binding affinity of wild-type influenza neuraminidase and its H274Y mutant to the antiviral drug zanamivir
Authors: Somasundaram B, Fee CJ, Fredericks R, Watson AJ, Fairbanks AJ.
Journal: J Mol Recognit (2015): 87
Susceptibility of human influenza A (H3N2) viruses to neuraminidase inhibitors isolated during 2011-2012 in China
Authors: Huang W, Tan M, Zhao X, Cheng Y, Li X, Guo J, Wei H, Xiao N, Wang Z, Wang D, Shu Y.
Journal: Zhonghua Yu Fang Yi Xue Za Zhi (2015): 481
Neuraminidase-inhibition assay for the identification of influenza A virus neuraminidase virus subtype or neuraminidase antibody specificity
Authors: Pedersen JC., undefined
Journal: Methods Mol Biol (2014): 27
Evaluation of the antigenic relatedness and cross-protective immunity of the neuraminidase between human influenza A (H1N1) virus and highly pathogenic avian influenza A (H5N1) virus
Authors: Lu X, Liu F, Zeng H, Sheu T, Achenbach JE, Veguilla V, Gubareva LV, Garten R, Smith C, Yang H, Stevens J, Xu X, Katz JM, Tumpey TM.
Journal: Virology (2014): 169
Partial antiviral activities detection of chicken Mx jointing with neuraminidase gene (NA) against Newcastle disease virus
Authors: Zhang Y, Fu D, Chen H, Zhang Z, Shi Q, Elsayed AK, Li B.
Journal: PLoS One (2013): e71688
Evaluation of the anti-neuraminidase antibodies in clinical trials of the live influenza vaccine of the A(H5N2) subtype
Authors: Smolonogina TA, Desheva Iu A, Rekstin AR, Mironov AN, Rudenko LG.
Journal: Vopr Virusol (2013): 31
Preparation and diagnostic utility of a hemagglutination inhibition test antigen derived from the baculovirus-expressed hemagglutinin-neuraminidase protein gene of Newcastle disease virus
Authors: Choi KS, Kye SJ, Jeon WJ, Park MJ, Kim S, Seul HJ, Kwon JH.
Journal: J Vet Sci (2013): 291
The fluorescence neuraminidase inhibition assay: a functional method for detection of influenza virus resistance to the neuraminidase inhibitors
Authors: Hurt AC, Okomo-Adhiambo M, Gubareva LV.
Journal: Methods Mol Biol (2012): 115