Amplite® Fluorimetric Neuraminidase Assay Kit *Blue Fluorescence*
Price | |
Catalog Number | |
Unit Size | |
Quantity |
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Overview | SDSProtocol |
Platform
Fluorescence microplate reader
Excitation | 320 nm |
Emission | 460 nm |
Cutoff | 420 nm |
Recommended plate | Solid black |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare Neuraminidase working solution (50 µL)
- Add Neuraminidase standards or test samples (50 µL)
- Incubate at 37°C or room temperature for 1 - 2 hours
- 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.
PREPARATION OF STOCK SOLUTION
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.
PREPARATION OF STANDARD SOLUTION
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.
PREPARATION OF WORKING SOLUTION
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.
SAMPLE EXPERIMENTAL PROTOCOL
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.
BL | BL | TS | TS |
NA1 | NA1 | ... | ... |
NA2 | NA2 | ... | ... |
NA3 | NA3 | ||
NA4 | NA4 | ||
NA5 | NA5 | ||
NA6 | NA6 | ||
NA7 | NA7 |
Table 2. Reagent composition for each well.
Well | Volume | Reagent |
NA1 - NA7 | 50 µL | Serial Dilution (0.312 to 20 mU/mL) |
BL | 50 µL | Blank Control |
TS | 50 µL | test sample |
- 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.
- 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.
- Incubate the reaction at 37°C or room temperature for 1 to 2 hours, protected from light. Note: 37°C incubation gives better results.
- Monitor the fluorescence increase at Ex/Em = 320/460 nm (Cutoff = 420 nm) with a fluorescence microplate reader.
Images
References
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
Authors: S, undefined and bulte MR, Gauger PC, Kitikoon P, Chen H, Perez DR, Roth JA, Vincent AL.
Journal: Vaccine (2016): 3773
Authors: Somasundaram B, Fee CJ, Fredericks R, Watson AJ, Fairbanks AJ.
Journal: J Mol Recognit (2015): 87
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
Authors: Pedersen JC., undefined
Journal: Methods Mol Biol (2014): 27
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
Authors: Zhang Y, Fu D, Chen H, Zhang Z, Shi Q, Elsayed AK, Li B.
Journal: PLoS One (2013): e71688
Authors: Smolonogina TA, Desheva Iu A, Rekstin AR, Mironov AN, Rudenko LG.
Journal: Vopr Virusol (2013): 31
Authors: Choi KS, Kye SJ, Jeon WJ, Park MJ, Kim S, Seul HJ, Kwon JH.
Journal: J Vet Sci (2013): 291
Authors: Hurt AC, Okomo-Adhiambo M, Gubareva LV.
Journal: Methods Mol Biol (2012): 115
Application notes
Acetylcholinesterase Inhibitory Activity of Pigment Echinochrome A
Ameliorative Effect of Novel Vitamin Formula with Herbal Extracts on Scopolamine-Induced Alzheimer's Disease
An Increase in Plasma Homovanillic Acid with Cocoa Extract Consumption Is Associated with the Alleviation of Depressive Symptoms in Overweight or Obese Adults
Attenuation of lysyl oxidase and collagen gene expression in keratoconus patient corneal epithelium corresponds to disease severity