At high pH, the hydroxyl groups of sugars become ionized, and the resulting negative charge can be exploited for separation on an anion exchange resin [21]. Neutral and amino sugars are eluted isocratically with 16 mM NaOH, and acidic sugars, such as uronic and sialic acids, are eluted with...
4 ) ∶n (gly) = 4∶1], NaOH[ n (OH - )∶n (gly) = 0129], H2O Phosphate buffer [17 ] 616 KH2PO4 [ n (H2PO - 4 ) ∶n (gly) = 4∶1], NaOH[ n (OH - )∶n (gly) = 1185], H2O 716 KH2PO4 [ n (H2PO ...
Digests were reconstituted in 1.4 ml of immunoaffinity purification buffer containing 50 mm MOPS, 10 mm Na2HPO4, and 50 mm NaCl, pH adjusted to 7.2 with NaOH. Resuspended peptides were cleared of any remaining precipitates with a 1-min 1,800 × g spin and placed on ice. One aliquot (...
5.8.6. Degradation Studies For degradation studies, three 4 mL vials containing 5 mg of PH-189 dissolved in 5 mL acetonitrile and 5 mL of 1 M of HCl, 1 M of NaOH, and 1 M of H2O2 was added to each vial. The samples were heated to 90 ◦C for 90 min and evaporated using ...
Staining was performed by using PnPP (Sigma, St Louis, MO) at a concentration of 0.6 mg/mL diluted in diethynolamine (DEA) buffer. Adding 2.4 M/L NaOH to the wells after 10 minutes at room temperature stopped the reaction. Absorption was measured at 405 nm. Extinctions obtained with ...
Uptake was terminated by washing the cells twice with ice-cold Hanks’ buffer, and cells were then lysed with 50 mM NaOH, and scintillation fluid (Ultima Gold, Packard Instruments) added. 3H-glycine uptake was quantified in counts per minute using a Unifilter-96 GF/C microplate scintillation ...
Dried samples were cocrystallized with sDHB matrix (5 mg/ml in 50% ACN, 1 mM NaOH). Spectra were recorded using the rapifleX mass spectrometer, and tandem MS/MS was performed for glycan identification. Data Preprocessing and Analysis The total ion current-normalized overall average spectrum of ...
Frozen cell pellets (107 cells) of mock or ST3GAL4 transfected MKN45 cells were directly resuspended in 2 mL of lysis buffer (50 mM Tris–HCl, 100 mM NaCl, 1 mM EDTA and protease inhibitor at pH 7.4) and stored on ice for 20 min. The cells were lysed using a Polytron homogenizer ...
IMAC purified fusion toxin was bound to the column in 27 mM KH2PO4, 46 mM NaOH, 10 mM EDTA, 0.75 M NaCl, 0.1% Tween 20, pH 8, and after washing in the same buffer, elution was performed in 0.1 M Na2CO3, 0.5 M NaCl, 0.1% Tween 20, at pH 11.3. The eluted sample was ...