Log In Sign up
The largest database of trusted experimental protocols

Sialidase

Manufactured by Merck Group
Visit Supplier
Sourced in United States

Sialidase is a laboratory enzyme that cleaves the glycosidic linkages of sialic acid residues from glycoproteins, glycolipids, and oligosaccharides. It functions to remove terminal sialic acid moieties from these molecules.

Automatically generated - may contain errors

Lab products found in correlation

Sialidase from clostridium perfringens, by Merck Group (2 mentions) Bicinchoninic acid assay, by Thermo Fisher Scientific (2 mentions) 96 well round bottom plate, by Corning (1 mentions) Fluorescein isothiocyanate (fitc), by Thermo Fisher Scientific (1 mentions) Cyanine cy3, by Jackson ImmunoResearch (1 mentions) Anti np, by Thermo Fisher Scientific (1 mentions) Secondary goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Secondary anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Pertussis toxin, by R&D Systems (1 mentions) Rpmi 1640, by R&D Systems (1 mentions) Maraviroc, by Bio-Techne (1 mentions) Bms ccr2 22, by Bio-Techne (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Gemini Bio (1 mentions) Specific antibody, by Thermo Fisher Scientific (1 mentions) Hrp conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Pvdf membrane, by Bio-Rad (1 mentions) Milli q water purification system, by Merck Group (1 mentions) Clostridium perfringens, by Merck Group (1 mentions) Cd41 percp cy5, by BioLegend (1 mentions)

23 protocols using sialidase

1

Siglec-F and Muc5b Western Blotting

Check if the same lab product or an alternative is used in the 5 most similar protocols
Expression levels of Siglec-F binding proteins and Muc5b were assayed by western blotting using standard separation and transfer techniques. For detection of Siglec-F binding proteins, the blots were incubated with Siglec-F-human IgG1 Fc chimera (Siglec-F-Fc, 0.5 μg/mL, R&D Systems, Minneapolis, MN), which was pre-complexed with ECL anti-human IgG, horseradish peroxidase-linked whole antibody (from sheep, 1:2500, GE Healthcare, UK), in blocking solution (4% bovine serum albumin in Tris-buffered saline containing 0.1% Tween-20) at room temperature for 1 hour. Anti-Muc5b goat polyclonal antibody I-12, used 1:300 from Santa Cruz Biotechnology, Santa Cruz, CA) was diluted in blocking solution. Binding of Siglec-F or antibody probes were detected by chemiluminescence using either the ECL western blotting detection system (GE Healthcare UK Ltd, England) or the SuperSignalWest Pico Chemiluminescent substrate (ThermoScientific). Siglec-F-Fc and antibody binding was quantified by scanning densitometry using NIH ImageJ software. Some samples were pretreated with sialidase (Clostridium perfringens, 1.6 mU/mL, 2 hr, 37°C, Sigma-Aldrich; or Vibrio cholera, 100 mU/mL, 1 hr, 37°C, kindly provided by Dr. Ronald Schnaar, Johns Hopkins University School of Medicine) to confirm sialidase sensitivity.
Kiwamoto T., Katoh T., Evans C.M., Janssen W.J., Brummet M.E., Hudson S.A., Zhu Z., Tiemeyer M, & Bochner B.S. (2014). Endogenous airway mucins carry glycans that bind Siglec-F and induce eosinophil apoptosis. The Journal of allergy and clinical immunology, 135(5), 1329-1340.e9.
+ Open protocol
+ Expand
2

Platelet Depletion and Desialylation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mice were injected intraperitoneally with 50 μl of rabbit anti-mouse platelet serum (RAMPS, Inter-cell Technologies) or intravenously by retroorbital injections with 50 mU of α2–3,6,8 neuraminidase (sialidase) from C. perfringens (Sigma-Aldrich).
Grozovsky R., Begonja A.J., Liu K., Visner G., Hartwig J.H., Falet H, & Hoffmeister K.M. (2014). The Ashwell-Morell receptor regulates hepatic thrombopoietin production via JAK2-STAT3 signaling. Nature medicine, 21(1), 47-54.
+ Open protocol
+ Expand
3

Sialidase Treatment of Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were recovered, washed twice with PBS, and suspended in phosphate buffer (pH 5.8). Sialidase (derived from Clostridium perfringens, Sigma) was added at a concentration of 100 mU/ml and reacted at 37 °C for 30 min.
Tian Y., Denda-Nagai K., Tsukui T., Ishii-Schrade K.B., Okada K., Nishizono Y., Matsuzaki K., Hafley M., Bresalier R.S, & Irimura T. (2022). Mucin 21 confers resistance to apoptosis in an O-glycosylation-dependent manner. Cell Death Discovery, 8, 194.
+ Open protocol
+ Expand
4

Modulation of BMDC Activation by Sialic Acid Analogs

Check if the same lab product or an alternative is used in the 5 most similar protocols
On day 0, day 3, and day 6 of the BMDC culture, 250 µM Ac53FaxNeu5Ac or DMSO was added to the medium, except for the dose titration experiment, where a concentration between 0 and 500 µM of Ac53FaxNeu5Ac was used. Sialidase treatment was performed with differentiated BMDCs on day 7 by adding 250 mU/ml Sialidase from Clostridium perfringens (Sigma-Aldrich) for 1 hour (h) at 37 °C. After incubation, the cells were washed thoroughly and used for experiments. For the TLR stimulation, 1*105 day 7 BMDCs were plated into 96-well round bottom plates (Corning) and stimulated with CpG (200 and 400 ng/ml) or LPS (1 and 10 ng/ml) for 18 h at 37 °C. After the stimulation, the cells were harvested and stained for flow cytometry analysis.
Balneger N., Cornelissen L.A., Wassink M., Moons S.J., Boltje T.J., Bar-Ephraim Y.E., Das K.K., Søndergaard J.N., Büll C, & Adema G.J. (2022). Sialic acid blockade in dendritic cells enhances CD8+ T cell responses by facilitating high-avidity interactions. Cellular and Molecular Life Sciences, 79(2), 98.
+ Open protocol
+ Expand
5

Viral Titer Quantification by Immunofluorescence

Check if the same lab product or an alternative is used in the 5 most similar protocols
Viral titers were determined by immunofluorescence (IF) on sub-confluent MDCKII cells as decribed12 (link). Rabbit polyclonal anti-H18 and secondary anti-rabbit IgG coupled to Cyanine Cy3 (Jackson ImmunoResearch, 1:500) antibodies were used to detect and quantify H18 positive cells.
Bat rectal swab samples were collected in 500 μL brain heart infusion (BHI) medium and subsequently filtered by adding additional 500 μL PBS through a 0.45 μm filter. The filtered flow-through was used for virus titration by performing tenfold dilution series in PBS. Viral titers were determined by IF on sub-confluent MDCKII cells pretreated with 100 mU per mL sialidase (Sigma) for 1 h and DEAE for 30 min at 37°C respectively, using rabbit polyclonal anti-NP (Thermo Fisher Scientific, 1:400) and the secondary goat anti-rabbit IgG coupled to FITC (Thermo Fisher Scientific, 1:400) antibodies.
Ciminski K., Ran W., Gorka M., Lee J., Malmlov A., Schinköthe J., Eckley M., Murrieta R.A., Aboellail T.A., Campbell C.L., Ebel G.D., Ma J., Pohlmann A., Franzke K., Ulrich R., Hoffmann D., García-Sastre A., Ma W., Schountz T., Beer M, & Schwemmle M. (2019). Bat influenza viruses transmit among bats but are poorly adapted to non-bat species. Nature microbiology, 4(12), 2298-2309.
+ Open protocol
+ Expand
6

Sialidase Treatment and Chemokine Receptor Blocking for CD8+ T Cell Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
For removing sialic acid residues, cell-sorted subsets of CD8+ T cells were treated with 0.1 units of sialidase (from Vibrio cholera, Sigma-Aldrich) in 1 ml RPMI 1640 (Life Technologies) containing 10% FBS (Gemini Bio-Products) and 2 mM L-glutamine for 2.5 hr at 37°C. For inhibiting Gi/o proteins, CD8+ T cells were pre-incubated with pertussis toxin (1 μg/ml (R and D Systems) in RPMI 1640 medium containing 10% FBS and 10 mM HEPES for 3 hr at 37°C. For blocking CCR2 and CCR5, pre-incubation was with BMS CCR2 22 (2 μM (Tocris, Minneapolis, MN) or Maraviroc (10 μM (Tocris), respectively, for 30 min at 37°C and inhibitors were left in the medium throughout the assay. For neutralizing CCL20, HUVEC monolayers in flow chambers were pre-treated for 2 hr at 37°C with 20 μg/ml anti-human CCL20/MIP-3α antibody (c67310; R and D Systems), and antibody was maintained at 10 ng/ml throughout the assay.
Lee C.H., Zhang H.H., Singh S.P., Koo L., Kabat J., Tsang H., Singh T.P, & Farber J.M. (2018). C/EBPδ drives interactions between human MAIT cells and endothelial cells that are important for extravasation. eLife, 7, e32532.
+ Open protocol
+ Expand
7

Influenza Virus Binding and Internalization

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
For influenza virus binding, A549 cells with FGFR1, FGFR4, or GFP overexpression were pretreated with or without 0.01 units of sialidase (Sigma-Aldrich) ml-1 at 37°C for 8 h [9 (link)], and then pre-cooled for 10 min, incubated with PR8 or H5N1 virus at a multiplicity of infection (MOI) of 1 at 4°C for 1 h, and finally washed twice with ice-cold PBS [1 (link)]. Cell lysates were prepared at 4°C with ice-cold RIPA buffer containing protease inhibitor cocktail. For virus internalization, cells with prebound virus were warmed to 37°C for 30 min and then washed with acidic cold PBS-HCl (pH 1.3) for detection of internalized virus particles, but not attached particles [9 (link)]. Cell lysates were obtained to determine influenza virus NP levels by Western blotting.
Liu X., Lai C., Wang K., Xing L., Yang P., Duan Q, & Wang X. (2015). A Functional Role of Fibroblast Growth Factor Receptor 1 (FGFR1) in the Suppression of Influenza A Virus Replication. PLoS ONE, 10(4), e0124651.
+ Open protocol
+ Expand
8

Far-Western Blot for Protein Interactions

Check if the same lab product or an alternative is used in the 5 most similar protocols
For Far-Western blot, vimentin or sialidase (Sigma) pretreated vimentin (for 0.5 h at 37°C) was separated by SDS-PAGE, then transferred to PVDF membranes (Bio-Rad) and blocked with 5% (w/v) skimmed milk for 2 h at 37°C. Subsequently, the membranes were washed thrice using TBST (TBS containing 0.01% Tween 20) and incubated with the recombinant fragments of SssP1 for 2 h at 37°C. After thrice wash, the processed membranes were incubated with the specific antibody (1:2000 dilution) (Thermo Fisher) for 1 h at 37°C. After another thrice wash, the membranes were stained with the HRP conjugated secondary antibodies (Thermo Fisher, 1:2000) for 1 h at 37°C. The positive bands were detected using the 3,3’-diaminobenzidine. Except for co-incubation with recombinant fragments of SssP1, the Western blot detecting ZO-1, occludin, claudin-5 and β-actin was performed in a similar procedure.
Pan Z., He P., Zhang Y., Gu Q., Chen S., Yu Y., Shao J., Wang K., Wu Z., Yao H, & Ma J. (2022). SssP1, a Fimbria-like component of Streptococcus suis, binds to the vimentin of host cells and contributes to bacterial meningitis. PLoS Pathogens, 18(7), e1010710.
+ Open protocol
+ Expand
9

Site-Specific Protein Glycosylation

Check if the same lab product or an alternative is used in the 5 most similar protocols
All solvents and chemicals were used without further purification, unless otherwise specified. The amino acids used in this study were purchased from Bachem. DMF (Dimethylformamide, HPLC grade), CH3CN and CH2Cl2 from Fisher, TFA (trifluoroacetic acid), DIEA (N,N-diisopropylethylamine), N,N′-Diisopropylcarbodiimide (DIC) and Boc-aminooxyacetic acid (Boc-Aoa-OH) and Sialidase (from Clostridium perfringens) were purchased from Sigma-Aldrich. DI-water (18 MΩ) was prepared using a Millipore Milli-Q water purification system. HCTU (o-(1-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) and MBHA resin (4-methylbenzhydrylamine, 0.65 mmol/g) were purchased from Peptides International. 1-Hydroxybenzotriazole (HOBT) was purchased from GL Biochem Shanghai. Rink amide (0.68 mmol/g) and Aminohexanoic acid (Ahx) were purchased from Novabiochem. TIS (Triisopropylsilane) was purchased from Oakwood Chemical. AmiconUltra Centrifugal filtration devices (0.5 mL, 50 MW cutoff) and TRITC-dye-conjugated Sambucus nigra (Elderberry Bark, SNA I) were respectively purchased from Millipore Inc. and EY Laboratories. Glycans 1a and 1b were obtained from chicken eggs as described by Kajihara, 12 (link)30 (link)44 and provided as a generous gift by GlyTech Inc. (Kyoto, Japan).
Palomo V., Cistrone P.A., Zhan N., Palui G., Mattoussi H, & Dawson P.E. (2018). Efficient Assembly of Quantum Dots with Homogenous Glycans Derived from Natural N-linked Glycoproteins. Bioconjugate chemistry, 29(9), 3144-3153.
+ Open protocol
+ Expand
10

Comprehensive Flow Cytometry Immunostaining Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
The following are primary reagents for flow cytometry and immunostaining unless specifically stated in individual method sections. These include directly conjugated primary antibodies to murine CD41-Percp-Cy5.5 (#133918), P-selectin-FITC (#304903, CD45-FITC (#103108), F4/80-PE (#123110), and Ly6G-PE (#127608), which were from BioLegend. Primary antibodies to murine CD31 (#ab119341) and F4/80 (#ab6640) were from Abcam. Secondary antibodies include goat anti-AH (#AF647, Abcam), and donkey anti-rat (#AF488, Abcam). Other reagents include Strep-Cy3 and Hoechst 33342 (#62249, Fisher Scientific), CellTracker Deep Red (#C34565 Life Technologies), and Zombie Aqua (#423101, BioLegend). Arthrobacter ureafaciens (#10269611001)- or Clostridium perfringens (#11586886001)-derived neuraminidase (sialidase) were purchased from Sigma.
Jiang Y., Tang Y., Hoover C., Kondo Y., Huang D., Restagno D., Shao B., Gao L., Michael McDaniel J., Zhou M., Silasi-Mansat R., McGee S., Jiang M., Bai X., Lupu F., Ruan C., Marth J.D., Wu D., Han Y, & Xia L. (2021). Kupffer cell receptor CLEC4F is important for the destruction of desialylated platelets in mice. Cell Death and Differentiation, 28(11), 3009-3021.
+ 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!