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α2 6 sialyllactose

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α2,6-sialyllactose is a carbohydrate compound that serves as a key component in various laboratory assays and analyses. It functions as a reference standard or building block for studying sialylated glycans and their interactions.

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Lab products found in correlation

α2 3 sialyllactose, by Merck Group (3 mentions) Neu clost, by Merck Group (2 mentions) Lactitol, by Merck Group (2 mentions) U0126, by Cell Signaling Technology (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) 24 well plate, by Thermo Fisher Scientific (2 mentions) Chelerythrine, by LC Laboratories (2 mentions) Absolute ethanol, by Merck Group (1 mentions) Fetuin from fetal calf serum, by Merck Group (1 mentions) Spectramax m5e microplate reader, by Molecular Devices (1 mentions) Na fluor influenza neuraminidase assay kit, by Thermo Fisher Scientific (1 mentions) 4 methylumbelliferone 4 mu, by Merck Group (1 mentions) Ammonium hydroxide, by Merck Group (1 mentions) Ammonium acetate, by Merck Group (1 mentions) Auto itc200, by Malvern Panalytical (1 mentions)

4 protocols using α2 6 sialyllactose

1

Characterizing CD22 Interactions by ITC

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ITC experiments were performed using an Auto-ITC200 (Malvern Instruments). ITC measurements of CD2220–330 WT and mutants (K23S, R120A/E, R131A/E/K/Q) with α2-6 sialyllactose were collected using 70–110 μM of CD22 in the cell and 0.65–1.18 mM of α2-6 sialyllactose (Sigma-Aldrich) in the syringe. A total of 16 injections were performed with an injection volume of 2.5 μl, injection duration of 5, and 180 s spacing between injections. The cell temperature was set to 25 °C, with a stirring speed of 750 r.p.m. and a filter period of 5 s. All experiments were repeated at least in duplicates, and values were averaged and standard errors were calculated, based on a near 1:1 restricted binding stoichiometry. For Fab binding to CD22, 5 μM of CD22 was placed in the cell and 50 μM Fab was present in the syringe. A total of 16 injections were performed with an injection volume of 2.5 μl, injection duration of 5 s, and 180 s spacing between injections. The cell temperature was set to 35 °C, with a stirring speed of 750 r.p.m. and a filter period of 5 s. All experiments were repeated in triplicate, and values were averaged and standard errors were calculated.
Ereño-Orbea J., Sicard T., Cui H., Mazhab-Jafari M.T., Benlekbir S., Guarné A., Rubinstein J.L, & Julien J.P. (2017). Molecular basis of human CD22 function and therapeutic targeting. Nature Communications, 8, 764.
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2

Trypanosoma cruzi Infection of HT1080 Cells

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HT1080 cells were maintained in DMEM with 3.7 g L−1 NaHCO3 and supplemented with 10% FBS (Gibco) at 37 °C in a humidified environment containing 5% CO2. Then, 5 × 104 cells per well (6 wells per treatment) were seeded in 24-well plates (Nunc). After 16 h, cells were infected with 5 × 105 culture-derived trypomastigotes of the Cvd strain or the clone Q501/3 for 16 h and washed. Media were replaced 48 hours pi with FBS free media and 24 h later conditioned supernatants were collected (referred as supernatants), centrifuged and frozen-stored at −80 °C. Alternatively, cells were treated with 1 μg mL−1 BSA, recombinant aTS, iTS or neuraminidases from Salmonella typhimurium (Neu Sal) (Biolabs) or Clostridium perfringes (Neu Clost) (Sigma) and were processed as the infected cells. Three independent experiments were performed for each strain and neuraminidases. aTS activity was confirmed in the presence of 10 mmol L−1 Lactitol (Sigma), α2,3-sialyllactose or α2,6-sialyllactose (Carbosynth) as previously described [17 (link)]. For signalling inhibition assays, cells were preincubated with 10 μmol L−1 of Chelerythrine (LC Laboratories), U0126 or PP2 (Cell Signalling).
Musikant D., Higa R., Rodríguez C.E., Edreira M.M., Campetella O., Jawerbaum A, & Leguizamón M.S. (2021). Sialic acid removal by trans-sialidase modulates MMP-2 activity during Trypanosoma cruzi infection. Biochimie, 186, 82-93.
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3

Sialic Acid Quantification Protocol

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Ammonium acetate, ammonium hydroxide, N-acetyl-D-neuraminic acid (C11H19NO9, MW 309.3 Da), α-2,3-sialyllactose, α-2,6-sialyllactose, and fetuin from fetal calf serum were obtained from Sigma-Aldrich (St. Louis, MO). N-Acetyl-D-[1,2,3-13C3]-neuraminic acid internal standard (13C3C8H19NO9, MW 312.3 Da) was acquired from Omicron Biochemicals, Inc. (South Bend, IN). All chemicals were of the highest purity available and used without further purification. NA-Fluor 2× Assay buffer, 66.6 mM 2-(N-morpholino) ethanesulfonic acid (MES) buffer, 8 mM CaCl2, pH 6.5, was from Applied Biosystems (Foster City, CA). The LC column was from Imtakt USA (Portland, OR), and influenza vaccines A–F (Table 1) were commercially available.
During the implementation of the fluorescence method, the NA-Fluor Influenza Neuraminidase Assay kit from Applied Biosystems, Cat No. 4457091, was used. Ethanol absolute, 4-methylumbelliferone (4-MU), and sodium chloride solution were from Sigma-Aldrich. 96-well opaque black flat-bottom plates from Corning (Corning, NY) were used with a SpectraMax M5e micro plate reader from Molecular Devices LLC (Sunnyvale, CA) for fluorescence measurements.
Solano M.I., Woolfitt A.R., Williams T.L., Pierce C.L., Gubareva L.V., Mishin V, & Barr J.R. (2017). Quantification of Influenza Neuraminidase Activity by Ultra-High Performance Liquid Chromatography and Isotope Dilution Mass Spectrometry. Analytical chemistry, 89(5), 3130-3137.
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4

Trypanosoma cruzi Infection of HT1080 Cells

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HT1080 cells were maintained in DMEM with 3.7 g L−1 NaHCO3 and supplemented with 10% FBS (Gibco) at 37 °C in a humidified environment containing 5% CO2. Then, 5 × 104 cells per well (6 wells per treatment) were seeded in 24-well plates (Nunc). After 16 h, cells were infected with 5 × 105 culture-derived trypomastigotes of the Cvd strain or the clone Q501/3 for 16 h and washed. Media were replaced 48 hours pi with FBS free media and 24 h later conditioned supernatants were collected (referred as supernatants), centrifuged and frozen-stored at −80 °C. Alternatively, cells were treated with 1 μg mL−1 BSA, recombinant aTS, iTS or neuraminidases from Salmonella typhimurium (Neu Sal) (Biolabs) or Clostridium perfringes (Neu Clost) (Sigma) and were processed as the infected cells. Three independent experiments were performed for each strain and neuraminidases. aTS activity was confirmed in the presence of 10 mmol L−1 Lactitol (Sigma), α2,3-sialyllactose or α2,6-sialyllactose (Carbosynth) as previously described [17 (link)]. For signalling inhibition assays, cells were preincubated with 10 μmol L−1 of Chelerythrine (LC Laboratories), U0126 or PP2 (Cell Signalling).
Musikant D., Higa R., Rodríguez C.E., Edreira M.M., Campetella O., Jawerbaum A, & Leguizamón M.S. (2021). Sialic acid removal by trans-sialidase modulates MMP-2 activity during Trypanosoma cruzi infection. Biochimie, 186, 82-93.
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