water-soluble biotin-dbco click chemistry tools cat #a116-5  (Click Chemistry Tools)

Journal: PLoS ONE

Article Title: Click-to-Capture: A method for enriching viable Staphylococcus aureus using bio-orthogonal labeling of surface proteins

doi: 10.1371/journal.pone.0234542

Figure Lengend Snippet: (A) Molecular structures of the metabolic labels used in this study; DALA = Azido-D-Alanine; LAHA = L-Azidohomoalanine; GLAZ = Azidogalactosamine; GNAZ = Azidoglucosamine; MNAZ = Azidomannosamine. (B) LAHA robustly labels S . aureus , as detected by direct cycloaddition of the fluorophore DBCO-MB-543 (DM543). MOCK = cells grown without exposure to metabolic label or DM543; CTRL = cells grown without metabolic label, but exposed to DM543 15 minutes; DALA, LAHA, GNAZ, GLAZ, MNAZ = cells exposed to 5 mM of the indicated metabolic label for 1 hour, followed by 15 minutes exposure to DM543. (C) Distribution of incorporated metabolic label, primarily at the equatorial plane of S . aureus . (D) LAHA labels S . aureus during log-phase (LOG, solid line) as well as stationary phase (STA, broken lines); CTRL = cells grown without metabolic label, but exposed to DM543 15 minutes (black lines); LAHA = cells grown with LAHA for 1 hour followed by 15 minute exposure to DM543 (yellow lines). n = 3 for each experiment; a representative example is shown for each. (E) Molecular structure of fluorophore dibenzocyclooctyne DBCO-MB-543 (DM543).

Article Snippet: The metabolic labeling agents Azido-homoalanine (LAHA), N-azidoacetylglucosamine tetracetylated (GNAZ), N-azidoacetylmannosamine tetracetylated (MNAZ), N-azidoacetylgalactosamine tetracetylated (GLAZ), DBCO-MB-543 octyne-fluorophore (DM543), and water-soluble DBCO-sulfo-biotin (DSB), were obtained from Click Chemistry Tools (Scottsdale, AZ).

Techniques:

Journal: PLoS ONE

Article Title: Click-to-Capture: A method for enriching viable Staphylococcus aureus using bio-orthogonal labeling of surface proteins

doi: 10.1371/journal.pone.0234542

Figure Lengend Snippet: (A) Structure of a water-soluble DBCO-sulfo-biotin (DSB), an octyne-biotin with a short, rigid linker, and structure of DBCO-PEGn-Biotin; the PEG linkers tested in this study ranged from 4–60 PEG repeats. (B) Short linker-length octyne-biotins support streptavidin binding to LAHA labeled cells. All cells were labeled for 1 hour with 5 mM LAHA, without (CTRL) or with the indicated octyne-biotin for 15 minutes, followed by streptavidin-alexafluor 546 (SAF546); DSB = DBCO-sulfo-biotin; DP4B = DBCO-PEG4-biotin; DP30B = DBCO-PEG30-biotin; DP60B = DBCO-PEG60-biotin. (C) DALA is accessible for click reaction, but not streptavidin binding. All cells were labeled for 1 hour with 5 mM DALA, without (CTRL) or with the indicated octyne-biotin for 15 minutes, followed by SAF546; a parallel DALA-labeled sample was incubated with the octyne-fluorophore DM543 to demonstrate the incorporation of DALA. Octyne-biotin designations are the same as those shown in panel A. n = 3 for each experiment; a representative example is shown.

Article Snippet: The metabolic labeling agents Azido-homoalanine (LAHA), N-azidoacetylglucosamine tetracetylated (GNAZ), N-azidoacetylmannosamine tetracetylated (MNAZ), N-azidoacetylgalactosamine tetracetylated (GLAZ), DBCO-MB-543 octyne-fluorophore (DM543), and water-soluble DBCO-sulfo-biotin (DSB), were obtained from Click Chemistry Tools (Scottsdale, AZ).

Techniques: Binding Assay, Labeling, Incubation

Journal: PLoS ONE

Article Title: Click-to-Capture: A method for enriching viable Staphylococcus aureus using bio-orthogonal labeling of surface proteins

doi: 10.1371/journal.pone.0234542

Figure Lengend Snippet: (A) S . aureus in whole blood (10 2 −10 4 CFU from a titered stock) was incubated with LAHA for 15 minutes, followed by exposure to DBCO-PEG4-Biotin (DP4B) or DMSO (CTRL). The labeled cells were incubated with 2.8 μm diameter Streptavidin M280 beads for 30 minutes with continuous agitation, and collected on a magnet; bead-bound and depleted fractions were tested for the presence of viable cells by plating compared to an input control sample; click-biotinylation allows for significant depletion of labeled cells relative to control (p < 0.01, t-test; n = 10). (B) Streptavidin bead diameter is a critical determinant of enrichment/depletion efficiency. Cells, grown as in panel A, were incubated with magnetic beads of varying diameters (see for comparative physical characteristics of all bead types), and the number of colony-forming units as a percent of the input was calculated by growth on solid media. DP4B (gold squares) = LAHA-labeled and DP4B biotinylated cells; CTRL (red circles) = LAHA-labeled control cells without biotinylation.

Article Snippet: The metabolic labeling agents Azido-homoalanine (LAHA), N-azidoacetylglucosamine tetracetylated (GNAZ), N-azidoacetylmannosamine tetracetylated (MNAZ), N-azidoacetylgalactosamine tetracetylated (GLAZ), DBCO-MB-543 octyne-fluorophore (DM543), and water-soluble DBCO-sulfo-biotin (DSB), were obtained from Click Chemistry Tools (Scottsdale, AZ).

Techniques: Incubation, Labeling, Control, Magnetic Beads