OPSS-PEG-NHS (molecular weight 5000) was purchased from Nanocs (New York, NY, USA). Trastuzumab (Herceptin) was a generous gift from Dr Virginia Borges (UC Denver, Denver, CO, USA). LLO from Escherichia coli transfected with the LLO-pEt29-DP-E3570 plasmid, kindly provided by Dr Dan Portnoy (UC Berkeley, Berkeley, CA, USA), was purified by the method described previously13 (link), 14 (link) and stored in storage buffer (50 mM phosphate buffer, pH 6.0, 1 M NaCl and 1 mM EDTA) without dithiothreitol to preserve its activity. Polylysine hydrobromide (molecular weight 37 000; degree of polymerization: 177) and 2-iminothiolane-HCl (Traut's reagent) were purchased from Sigma Life Science (St Louis, MO, USA). CL-4B Sepharose used for the purification of the one-component complexes was purchased from Amersham Biosciences (Uppsala, Sweden). All other reagents, unless otherwise specified, were purchased from Thermo Fisher Scientific (Pittsburgh, PA, USA).
Polylysine hydrobromide
Manufactured by Merck Group
Sourced in United States
Polylysine hydrobromide is a synthetic amino acid polymer. It is a white, hygroscopic powder that is soluble in water. The primary function of polylysine hydrobromide is as a laboratory reagent, commonly used in various biochemical and cell culture applications.
Automatically generated - may contain errors
4 protocols using polylysine hydrobromide
1
Trastuzumab-LLO Conjugation Protocol
2
Polyelectrolyte Solutions for Biomaterials
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Poly(acrylic acid sodium salt) (PAA), poly(diallyldimethylammonium
chloride) (PDDA), poly-Lysine hydrobromide (poly-Lys), poly(ethylenimine)
(PEI), poly-threonine (poly-Thr), and poly-glutamic acid sodium salt
(poly-Glu) were purchased from Sigma. The polymers were dissolved
at 2 mg/mL in 20 mM phosphate buffer with 200 μM EDTA, and the
pH was set to 8.
chloride) (PDDA), poly-Lysine hydrobromide (poly-Lys), poly(ethylenimine)
(PEI), poly-threonine (poly-Thr), and poly-glutamic acid sodium salt
(poly-Glu) were purchased from Sigma. The polymers were dissolved
at 2 mg/mL in 20 mM phosphate buffer with 200 μM EDTA, and the
pH was set to 8.
3
P. aeruginosa Biofilm Formation Assay
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P. aeruginosa strain PA01 in 20 mL of Mueller-Hinton II broth cation adjusted (Becton Dickinson Diagnostic Systems, Inc., Sparks, MD, USA) was grown overnight at 37°C with continuous shaking at 250 rpm. The culture was then adjusted to a concentration of 1–5×108 cfu/mL, and 200 µL was used to inoculate µ-Slide 8 glass bottom wells (Ibidi, cat. num. 80827, Munich, Germany). Each well was previously coated with a 0.01% (w/v) poly-lysine hydrobromide (Sigma-Aldrich, Dorset, UK) solution to enhance bacterial cell adhesion and to prevent biofilm removal during the experiments. The slides were incubated at 37°C for 24 h to allow biofilm formation.
4
Trastuzumab-Listeriolysin O Conjugate Synthesis
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