The quantification of biotinylation was carried out by using a Quant Tag™ Biotin Kit (Vector Laboratories, Burlingame, CA). Samples were tested in triplicate against a known biotin standard curve to determine the number of biotins per allergen molecule.
Zeba desalt spin column
Zeba desalt spin columns are laboratory equipment designed to remove salts, buffers, and other small molecules from protein samples. They work by using a size-exclusion resin to selectively retain the protein of interest while allowing smaller molecules to pass through. This process is known as buffer exchange or desalting.
Lab products found in correlation
48 protocols using zeba desalt spin column
Biotinylation of Allergen Proteins
The quantification of biotinylation was carried out by using a Quant Tag™ Biotin Kit (Vector Laboratories, Burlingame, CA). Samples were tested in triplicate against a known biotin standard curve to determine the number of biotins per allergen molecule.
Biotinylation and Functionalization of Antibodies
For mAb biotinylation, 100 g of antibodies were diluted in 400 L of 0.1 M borate buffer pH 9.0 containing 6 L of biotin (Sigma-Aldrich) in DMF at 1 mg/mL and incubated for 30 min at room temperature. Then, 100 L of 1 M Tris HCl buffer pH 8.0 were added and incubated for 15 min. Finally, the biotinylated mAb was purified from free biotin on Zeba Desalt Spin column (Thermo Scientific) in 0.1 M potassium phosphate buffer pH 7.4 with 0.15 M NaCl. The absorbance of the final solution was measured between 280 and 320 nm to determine the concentration of the purified biotinylated antibody. biotinylated antibodies were then mixed at room temperature with streptavidin coated beads for 30 min, washed four times in PBS 0.1% BSA and stored in PBS 0.1% BSA at 4 C until use.
Fluorescent Antibody Conjugation Protocol
CD Spectroscopy Analysis of Protein Redox State
Fluorescent Diabody Conjugation Protocol
Biotinylation of Antibodies and IgG
Fluorescent Labeling of Protein Nanoparticles
Preparation of DDM-Solubilized Membrane Proteins
Japan). UQ-10, dithiothreitol (DTT), and trans-4,5-dihydroxy-1,2-dithiane
(oxidized DTT) were purchased from Wako Pure Chemical Industries Ltd.
(Osaka, Japan). UQ-1 was purchased from Sigma-Aldrich (St. Louis,
MO). NeutrAvidin, Zeba Desalt Spin Column, and tris(2-carboxyethyl)phosphine
(TCEP) were obtained from Thermo Fisher Scientific Inc. (Yokohama,
Japan). Maleimide–poly(ethylene glycol) (malPEG, SUNBRIGHT
ME-050MA, 5 kDa) was purchased from NOF Corporation (Tokyo, Japan).
Unless stated otherwise, all other reagents were purchased from Nacalai
Tesque Co. (Kyoto, Japan). Type 1 ultrapure water (MilliQ water, Merck
Ltd., Tokyo, Japan) was used in all experiments.
Quantitative Analysis of RhoA Dynamics
Quantitative Anti-drug Antibody Assay
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