Log In Sign up
The largest database of trusted experimental protocols

Na star influenza neuraminidase inhibitor resistance detection kit

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in United States

The NA-Star Influenza Neuraminidase Inhibitor Resistance Detection Kit is a laboratory tool designed to detect the presence of influenza neuraminidase inhibitor resistance. The kit provides a standardized method for analyzing influenza virus samples and determining their resistance to certain antiviral medications.

Automatically generated - may contain errors

Lab products found in correlation

Prism 7, by GraphPad (2 mentions) Microtiter plate reader, by Agilent Technologies (2 mentions) Prism software, by GraphPad (2 mentions) Synergy h1, by Agilent Technologies (1 mentions) Infinite 200 pro, by Tecan (1 mentions) Victor 1420 multilabel counter, by PerkinElmer (1 mentions) Glomax multi detection system, by Promega (1 mentions) Varioskan flash, by Thermo Fisher Scientific (1 mentions) Prism 8, by GraphPad (1 mentions)

Spelling variants of this product

NA-Star kit (7 citations) NA-Star™ Influenza NA Inhibitor Resistance Detection kit (2 citations) Influenza neuraminidase kit (2 citations) Resistance Detection Kit (2 citations) NA-STAR (2 citations)

18 protocols using na star influenza neuraminidase inhibitor resistance detection kit

1

Enzymatic Activity Measurement of Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
To measure the enzymatic activities of the respective proteins, an NA-Star assay was performed using the NA-Star influenza neuraminidase inhibitor resistance detection kit (ThermoFisher) as per the manufacturer’s instructions. In this assay, the proteins were diluted to a starting concentration of 10 μg/mL and then serially diluted 1:3 across the plate. The Mich15 N1-VASP protein was used as a positive control. The signal was based on the luminescence readout and measured using a Synergy H1 hybrid multimode microplate reader (BioTek). The data were analyzed using GraphPad Prism 7.
Strohmeier S., Amanat F., Zhu X., McMahon M., Deming M.E., Pasetti M.F., Neuzil K.M., Wilson I.A, & Krammer F. (2021). A Novel Recombinant Influenza Virus Neuraminidase Vaccine Candidate Stabilized by a Measles Virus Phosphoprotein Tetramerization Domain Provides Robust Protection from Virus Challenge in the Mouse Model. mBio, 12(6), e02241-21.
+ Open protocol
+ Expand
2

Neuraminidase Activity Comparison

Check if the same lab product or an alternative is used in the 5 most similar protocols
The rEgH9N2(P), rEgH9N2(P)-av370L, and rEgH9N2(P)-av400L strains were diluted into 128 (27) HAU at 4 °C to match the amount of virus in the absence of neuraminidase activity and used to compare the neuraminidase activities of each virus using the NA-Star™ Influenza Neuraminidase Inhibitor Resistance Detection Kit according to the manufacturer’s instructions (Thermo Fisher Scientific). Each virus was diluted five-fold with NA-Star™ assay buffer, and 50 μL of each dilution was transferred to a 96-well white plate in triplicate. The NA-Star™ substrate diluted 1:1000 with the assay buffer was added to every well and reacted at room temperature for 30 min. After the addition of 60 μL of the NA-Star™ accelerator and shaking for 2 s, luminescence was measured by Infinite 200 PRO (TECAN).
An S.H., Son S.E., Song J.H., Hong S.M., Lee C.Y., Lee N.H., Jeong Y.J., Choi J.G., Lee Y.J., Kang H.M., Choi K.S, & Kwon H.J. (2022). Selection of an Optimal Recombinant Egyptian H9N2 Avian Influenza Vaccine Strain for Poultry with High Antigenicity and Safety. Vaccines, 10(2), 162.
+ Open protocol
+ Expand
3

Neuraminidase Inhibition Assay for VLPs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Purified VLPs preparations were tested for enzymatic neuraminidase activity using the NA-Star Influenza Neuraminidase Inhibitor Resistance Detection Kit (ThermoFisher Scientific, Waltham, MA, USA) according to the manufacturer’s instructions. Briefly, VLP solutions were diluted in NA-Star Assay Buffer and then incubated with substrate for 30 min at room temperature. NA-Star accelerator solution was then injected followed by measurement of the chemiluminescent signal in relative luminescent units (RLU) via Modulus II Microplate Luminometer (Promega, Madison, WI, USA). Inhibition of neuraminidase activity was measured similarly, with collected sera diluted in NA-Star Assay Buffer followed by addition of 40 ng or 25 ng of NA2 or NA1 VLPs, respectively. Sera and VLPS were incubated at 37°C for 30 minutes followed by addition of substrate and incubation at room temperature for 30 minutes. Plates were read after addition of accelerator using the Modulus II Microplate Luminometer. The highest sera dilution inhibiting at least 50% of NA activity was reported as the neuraminidase inhibition (NAI) titer.
Menne Z., Pliasas V.C., Compans R.W., Glover S., Kyriakis C.S, & Skountzou I. (2021). Bivalent vaccination with NA1 and NA2 neuraminidase virus-like particles is protective against challenge with H1N1 and H3N2 influenza A viruses in a murine model. Virology, 562, 197-208.
+ Open protocol
+ Expand
4

Influenza Neuraminidase Inhibitor Resistance Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols
The NA enzymatic activity was determined by using the NA-Star™ Influenza Neuraminidase Inhibitor Resistance Detection Kit (ThermoFisher). Briefly, the recombinant N1 mutant proteins were diluted to a starting concentration of 10 µg/mL then serially diluted (1:3). The assay was performed according to the manufacturer’s instructions. As a positive control, recombinant A/Michigan/45/2015 N1 containing the VASP tetramerization domain was used.
Strohmeier S., Carreño J.M., Brito R.N, & Krammer F. (2021). Introduction of Cysteines in the Stalk Domain of Recombinant Influenza Virus N1 Neuraminidase Enhances Protein Stability and Immunogenicity in Mice. Vaccines, 9(4), 404.
+ Open protocol
+ Expand
5

Influenza Neuraminidase Inhibition Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
The NA-Star Influenza Neuraminidase Inhibitor Resistance Detection Kit (Applied Biosystems) was used to measure the ability of the anti-NA mAbs to inhibit the ability of viral NA to cleave a small, soluble chemiluminescent substrate. The assay was performed according to the manufacturer’s protocol. In brief, 25 μl of four-fold diluted antibodies (50–0.01 μg/ml) was transferred to a white, flat bottom 96-well plate. Then, 25 μl of NA-displaying VLPs was added to each well and the plate was shaken and incubated for 20 min at 37 °C. The NA-Star substrate was prepared shortly before use and 10 μl of the substrate was added to all wells. The plate was then incubated for 30 min at room temperature, and 60 μl of NA-Star accelerator solution was added to all wells immediately before the plate was read by using a microtiter plate reader (Turner BioSystems). The IC50 value was determined by nonlinear regression analysis (GraphPad Prism software).
Yasuhara A., Yamayoshi S., Kiso M., Sakai-Tagawa Y., Okuda M, & Kawaoka Y. (2022). A broadly protective human monoclonal antibody targeting the sialidase activity of influenza A and B virus neuraminidases. Nature Communications, 13, 6602.
+ Open protocol
+ Expand
6

Neuraminidase Inhibitory Activity Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Serum samples from NA1 and NA2 immunized and naive mice were assayed for NA enzymatic inhibition using the NA-Star influenza neuraminidase inhibitor resistance detection kit (Applied Biosystems). To measure sera-mediated inhibition, immunized and naive sera was serially diluted two-fold in NA-Star assay buffer in white, flat-bottom, 96-well cell culture plates. Virus (B/Florida/04/06 or B/Malaysia/2506/04) was diluted to the determined 3EC50 (half-maximum effective concentration) and 25 μL was added to each well. The plates were incubated for 30 min at 37 °C. Data points were expressed as percent inhibition of maximal NA enzymatic activity, which was determined by the activity of virus without the addition of sera. ELISA signals were fit with an inhibition regression algorithm and IC50 values determined using GraphPad Prism.
Zeigler D.F., Gage E, & Clegg C.H. (2021). Epitope-targeting platform for broadly protective influenza vaccines. PLoS ONE, 16(5), e0252170.
+ Open protocol
+ Expand
7

Influenza Neuraminidase Inhibitor Resistance

Check if the same lab product or an alternative is used in the 5 most similar protocols
The susceptibility of H7N9 viruses to oseltamivir carboxylate, zanamivir and peramivir was determined by the NA-Star™ Influenza Neuraminidase Inhibitor Resistance Detection Kit (Applied Biosystems, Foster City, CA, USA) according to the manufacturer's instructions. The chemiluminescent signal was measured with a Victor 1420 multi-label counter (PerkinElmer, Waltham, MA, USA).
Zhang X., Song Z., He J., Yen H.L., Li J., Zhu Z., Tian D., Wang W., Xu L., Guan W., Liu Y., Wang S., Shi B., Zhang W., Qin B., Cai J., Wan Y., Xu C., Ren X., Chen H., Liu L., Yang Y., Zhou X., Zhou W., Xu J., Zhang X., Peiris M., Hu Y, & Yuan Z. (2014). Drug susceptibility profile and pathogenicity of H7N9 influenza virus (Anhui1 lineage) with R292K substitution. Emerging Microbes & Infections, 3(11), e78-.
+ Open protocol
+ Expand
8

Measuring H6N1 Neuraminidase Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols
The neuraminidase activity of H6N1 virus was determined using the NA-Star Influenza Neuraminidase Inhibitor Resistance Detection Kit (Applied Biosystems, Foster city, California, USA) according to the manufacturer’s protocol. This assay used a chemiluminescent substrate to measure the activity of neuraminidases. In brief, H6N1 viruses were incubated with various concentrations of PSE for 10 min at 37 °C, and then the neuraminidase substrate, 4-MU-NANA [2-(4-methylumbelliferyl)-A-d-nacetylneuraminic acid sodium] was added and incubated for 2 h in the dark. The luminescence signal indicating neuraminidase activity was determined with fluorescence spectrophotometer (excitation 365 nm, emission 460 nm).
Yang C.H., Tan D.H., Hsu W.L., Jong T.T., Wen C.L., Hsu S.L, & Chang P.C. (2014). Anti-influenza virus activity of the ethanolic extract from Peperomia sui. Journal of Ethnopharmacology, 155(1), 320-325.
+ Open protocol
+ Expand
9

Quantifying Influenza Neuraminidase Inhibition

Check if the same lab product or an alternative is used in the 5 most similar protocols
The NA-Star® Influenza Neuraminidase Inhibitor Resistance Detection Kit (Applied Biosystems) was used to quantify the inhibition of NA activity (cleavage of a small chemiluminescent substrate) in the presence of NA-mAbs. The experiments were performed according to the manufacturer’s protocol. Briefly, mAbs were diluted in NA-Star Assay Buffer to a concentration of 100 μg/mL, and subsequently serially diluted 1:3. Twenty-five microliters from each dilution were transferred to a white, flat bottom 96-well cell culture plate and mixed with 25 μL/well of B/Phuket/3073/2013 virus at a predetermined 2 × EC50 for 20 min at 37°C. NA-Star Substrate (10 μL/well) was added after the incubation, and the plates were incubated at RT for 30 min. NA-Star accelerator solution (60 μL/well) was added to the plates immediately before the readout. The chemiluminescent signal was detected by a microtiter plate reader (Bio-Tek) and analyzed using Microsoft Excel and GraphPad Prism 7.
Madsen A., Dai Y.N., McMahon M., Schmitz A.J., Turner J.S., Tan J., Lei T., Alsoussi W.B., Strohmeier S., Amor M., Mohammed B.M., Mudd P.A., Simon V., Cox R.J., Fremont D.H., Krammer F, & Ellebedy A.H. (2020). Human Antibodies Targeting Influenza B Virus Neuraminidase Active Site Are Broadly Protective. Immunity, 53(4), 852-863.e7.
+ Open protocol
+ Expand
10

Influenza Neuraminidase Inhibitor Resistance

Check if the same lab product or an alternative is used in the 5 most similar protocols
The NA-Star influenza neuraminidase inhibitor resistance detection kit (Applied Biosystems) was used and manufacturer's instructions were followed. Briefly, 25 μl of each antibody in serial two-fold dilutions in NA-Star assay buffer (starting concentration 100 μg/ml) was mixed with 25 μl of virus at the determined 4× EC50 (half maximum effective concentration) and incubated at 37°C for 20 min. After adding 10 μl of 1000× diluted NA-Star substrate, the plates were incubated for 30 min. Reaction was stopped by adding 60 μl of NA-Star accelerator. The chemiluminescence was determined using the DTX 880 plate reader. IC50 values were determined using Prism software. The assay was performed in duplicate 2-3 times.
Henry C., Zheng N.Y., Huang M., Cabanov A., Thatcher Rojas K., Kaur K., Andrews S.F., Palm A.K., Chen Y.Q., Li Y., Hoskova K., Utset H.A., Vieira M.C., Wrammert J., Ahmed R., Holden-Wiltse J., Topham D.J., Treanor J.J., Ertl H.C., Schmader K.E., Cobey S., Krammer F., Hensley S.E., Greenberg H., He X.S, & Wilson P.C. (2019). Influenza virus vaccination elicits poorly adapted B cell responses in elderly individuals. Cell host & microbe, 25(3), 357-366.e6.
+ 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!