Ultrapure urea and guanidinium chloride were purchased from MP Biomedicals (CA, USA). All other reagents were purchased from Sigma Chemical Co. (MO, USA) or ThermoFisher (MA, USA). The C-amidated, N-acetylated AWPAK peptide was synthesised by ChinaPeptides (Shanghai, China). SOD1 H43Y noloops and Escherichia coli cytochrome b562 F65W proteins were donated by Mikael Oliveberg (University of Stockholm) and Paul Barker (Cambridge University), respectively. α-Spectrin expression vectors were gifts from Jane Clarke (Cambridge University). LQRRRETQV and LQRRRETQ-Abu peptides and the PDZ1 expression vector were gifts from Per Jemth (Uppsala University). Barley chymotrypsin inhibitor-2 (CI2), murine FBP28 WW domain, hepatitis B core protein, the PDZ1 of the post-synaptic density protein 95 (PSD-95) with the F95W mutation (PDZ1 F95W) and α-spectrin constructs were expressed and purified as reported [10,16,33,34] . Protein concentrations were determined spectrophotometrically from calculated molar extinction coefficients.
Ultrapure urea
Manufactured by MP Biomedicals
Sourced in United States
Ultrapure urea is a laboratory reagent used as a high-purity chemical compound. It is a crystalline solid form of the organic compound urea, with a chemical formula of CO(NH2)2. Ultrapure urea is designed to meet the stringent purity requirements of various scientific and analytical applications.
Automatically generated - may contain errors
Lab products found in correlation
Guanidinium chloride, by MP Biomedicals (1 mentions)
Refracto 30px, by Mettler Toledo (1 mentions)
Urea, by MP Biomedicals (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
Sodium acetate, by Thermo Fisher Scientific (1 mentions)
Hplc grade methanol and water, by Thermo Fisher Scientific (1 mentions)
Trifluoroacetic acid, by Merck Group (1 mentions)
3 protocols using ultrapure urea
1
Protein Expression and Purification
2
Denaturation Kinetics of SOD1 Variants
Check if the same lab product or an alternative is used in the 5 most similar protocols
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All experiments were
performed at 298 K, with 10 mM MES (pH 6.4) as background buffer,
unless otherwise stated. Stock solutions of 2 M NaAc (pH 6.3) (Millipore
Sigma) buffer and 2 M NaCl (VWR) solutions were prepared according
to standard practices. During preparation of the 2 M polyacetate stock
solutions, special consideration had to be taken to reach an ionic
strength that was comparable with the salt solutions, as well as a
stable pH. The preparation protocol has been extensively described
in theSupporting Information .
To
avoid excessive screening of charge interactions, as, e.g., observed
for GdmCl, we opted here for Ultrapure urea (MP Biomedicals) as the
denaturation agent; 5–9 M urea was used for the SOD1barrel unfolding kinetic data, while 5 M urea was used to denature the
protein in preparation for the refolding kinetics. When the U state
was studied with NMR, 4 M urea and 5 M urea was used as denaturing
conditions for SOD1I35A and SOD1barrel, respectively.
Before use, urea was prepared fresh and had its concentration verified
using the refractive index22 (link) with a Refracto
30PX portable refractometer (Mettler Toledo).
performed at 298 K, with 10 mM MES (pH 6.4) as background buffer,
unless otherwise stated. Stock solutions of 2 M NaAc (pH 6.3) (Millipore
Sigma) buffer and 2 M NaCl (VWR) solutions were prepared according
to standard practices. During preparation of the 2 M polyacetate stock
solutions, special consideration had to be taken to reach an ionic
strength that was comparable with the salt solutions, as well as a
stable pH. The preparation protocol has been extensively described
in the
To
avoid excessive screening of charge interactions, as, e.g., observed
for GdmCl, we opted here for Ultrapure urea (MP Biomedicals) as the
denaturation agent; 5–9 M urea was used for the SOD1barrel unfolding kinetic data, while 5 M urea was used to denature the
protein in preparation for the refolding kinetics. When the U state
was studied with NMR, 4 M urea and 5 M urea was used as denaturing
conditions for SOD1I35A and SOD1barrel, respectively.
Before use, urea was prepared fresh and had its concentration verified
using the refractive index22 (link) with a Refracto
30PX portable refractometer (Mettler Toledo).
3
Preparation of Apolipoprotein Mb Variants
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