Mpeg maleimide

Manufactured by Merck Group

Sourced in United States

MPEG-maleimide is a chemical compound used in various laboratory applications. It consists of a methoxy-polyethylene glycol (MPEG) molecule with a maleimide functional group attached. The maleimide group allows for the selective conjugation of the MPEG molecule to thiol-containing compounds, making it a useful tool in protein modification and other biomolecular research.

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2 protocols using mpeg maleimide

Efficient Cysteine Labeling of Nanobodies

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Immediately before labelling, the Nbs were transferred into nonreducing buffer (10 mM HEPES pH 7.6, 150 mM NaCl) using a PD-10 desalting column. Nbs were immediately mixed with a fivefold molar excess of para-chloromercuribenzoic acid (PCMB) or a tenfold molar excess of mercury(II) acetate and incubated on ice for 1 h. PCMB labelling was quenched after 1 h by the addition of a 100-fold molar excess of iodoacetamide. To quantify the PCMB labelling efficiency, the remaining free cysteines were reacted with a tenfold molar excess of MPEG–maleimide (Sigma–Aldrich, catalogue No. 99126-64-4) on ice for 1 h. The samples were analysed by nonreducing SDS–PAGE and the ratio of Nb that had reacted with MPEG–maleimide to Nb that had reacted with PCMB was compared and quantified using ImageJ (Hartig, 2013 ▸ ).

Hansen S.B., Laursen N.S., Andersen G.R, & Andersen K.R. (2017). Introducing site-specific cysteines into nanobodies for mercury labelling allows de novo phasing of their crystal structures. Acta Crystallographica. Section D, Structural Biology, 73(Pt 10), 804-813.

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Synthesis and Characterization of S-Nitrosoglutathione

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3-(Trimethoxysilyl) propyl methacrylate, methacrylic acid (MAA), ethylene dimethacrylate (EDMA), toluene, dodecanol, and azobis-(isobutyronitrile) (AIBN) were purchased from Sigma-Aldrich (MO, USA). Fused-silica capillaries with 530 μm i.d. × 720 μm o.d. were obtained from Polymicro Technologies™. Acetonitrile (ACN), methanol, acetone and formic acid of LC grade were also from Sigma-Aldrich (MO, USA). Ultra-Pure water was obtained using an in-house purification system. Mercury chloride (HgCl₂), ethylenediaminetetraacetic acid (EDTA), potassium dihydrogen phosphate (KH₂PO₄), mPEG-maleimide, sodium nitrite, glutathione (GSH) and ¹³C₂,¹⁵N-labeled G*SH were from Sigma-Aldrich (MO, USA) and were used without additional purification. Vivaspin 3000 MWCO membrane filters were from Sartorius Stedim (NA, USA). All sample preparations were carried out in the dark at 4°C unless otherwise stated. GSNO, ¹³C₂,¹⁵N-labeled G*SNO and 2-[1-(dimethylamino)ethanethioate]triphenylphosphine (derivatizing reagent) were synthesized as described earlier,^{18 (link)} and were confirmed by LC/ESI-HR-MS (see Figure S-1 and Supporting Information for details).

Wang X., Garcia C.T., Gong G., Wishnok J.S, & Tannenbaum S.R. (2018). Automated Online Solid Phase Derivatization for Sensitive Quantification of Endogenous S-Nitrosoglutathione and Rapid Capture of Other Low-Molecular-Mass S-Nitrosothiols. Analytical chemistry, 90(3), 1967-1975.

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