Quantitative densitometry from the point of view of information theory

Pardowitz, Iancu; Ehrhardt, Wolfgang; Neuhoff, Volker

doi:10.1002/elps.1150110509

Cited by 8 publications

(3 citation statements)

References 13 publications

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“…Then the entire gel was electrophoresed and stained with coomassie brilliant blue to visualize location of proteins and determine if they were immobilized into polyacrylamide hydrogels in the well or free to migrate through the gel. The amounts of immobilized protein and migrated protein were quantified by gel densitometry to determine the percentage of protein immobilized in the sample wells. , We tested this method on the pentenyl-Protein G first, but unfortunately, the pentenyl-protein G migrated into the gel during electrophoresis just as the unmodified Protein G and only a negligible amount of pentenyl-Protein G (4.5%) was immobilized into the hydrogel in the well (Figure S1). This suggests that the pentenyl-Protein G could not be immobilized by free radical copolymerization using APS as the initiator because of the inertness of the α-olefin structure of 4-pentenyl group .…”

Section: Resultsmentioning

confidence: 99%

Synthesis of Polymerizable Protein Monomers for Protein-Acrylamide Hydrogel Formation

Xiao

Tolbert

2009

Biomacromolecules

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A novel method to produce protein polymer conjugates for protein-acrylamide hydrogel formation is described. Alkenes are incorporated onto the N-terminus of expressed proteins to produce polymerizable protein monomers that can be utilized in protein-acrylamide copolymerization. A 4-vinylbenzoic acid thioester was synthesized and attached to the N-termini of two protein models, the immunoglobulin-binding protein Protein G and the bacterial enzyme xanthine-guanine phosphoribosyltransferase (GPRT), utilizing native chemical ligation. N-terminal cysteine containing proteins utilized in native chemical ligation reactions were generated from His-tagged fusion proteins using tobacco etch virus NIa (TEV) protease cleavage. The 4-vinylbenzyl functionalized proteins were good substrates for immobilizing proteins into polyacrylamide hydrogels via free radical induced protein-acrylamide copolymerization. The protein copolymerization procedures developed in this report are mild enough to allow proteins to retain measurable biological activity as demonstrated by the retention of immunoglobulin binding ability by immobilized Protein G and enzymatic activity of immobilized GPRT.

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Section: Resultsmentioning

confidence: 99%

Synthesis of Polymerizable Protein Monomers for Protein-Acrylamide Hydrogel Formation

Xiao

Tolbert

2009

Biomacromolecules

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“…The combination of scanning photometry and image analysis is a sensitive, versatile, and accurate method for the evaluation of slab gels [49]. The process of densitometric evaluation of gels can be described as a series of message channels connected one behind the other, allowing for the basic principles of information theory to be applied [50].…”

Section: Two-dimensional Microslab Gel Electrophoresismentioning

confidence: 99%

Microelectrophoresis and auxilary micromethods

Neuhoff

2000

Electrophoresis

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In this retrospect of approximately 30 years of work with micromethods, some of them developed in our own laboratory, their principles and application to different separation problems are described, such as one‐ and two‐dimensional microelectrophoresis in capillaries and microslab gels, isoelectric focusing in capillaries or microslab gels, microchromatography, microphotometry, and microfluorometry for qualitative and quantitative evaluation of separation patterns. In addition, some useful auxiliary methods are also described, e.g., a method for quantitative protein determination in a microliter volume when neither the volume nor the protein content in that volume are known, and methods for the determination of glycoproteins, amino acids, and sugars in the picomole range.

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“…The signals are independent of one another as ifthey were transmitted randomly with given probabilities p l . An observer receives a certain amount of information from each signal generated, for which the mean value of the information is, according to Shannon H = -C 2 pi log(pi) (1) i Since the probabilityp, always lies between 0 and 1, the mean information H always has a positive valuie. In this definition, C Correspondence: Dr.…”

Section: Introductionmentioning

confidence: 99%