Theoretical and practical aspects of glutaraldehyde fixation

Hopwood, David A.

doi:10.1007/978-1-4899-3260-0_2

Cited by 213 publications

(71 citation statements)

References 115 publications

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“…It has been shown that binding of recombinant sAPP (sAPPrec) is highly sensitive to aldehyde fixation. Because of the preferential reactions of aldehyde groups with lysine residues (Hopwood 1972), this inhibitory effect on sAPPrec binding points to the proteinaceous nature of sAPPrec binding sites (Hoffmann et al 1999). Apparently, the number of binding sites for sAPPrec is not constant because it appears to be dependent, for example in the neuronal cell line B 104, on the state of differentiation (Hoffmann et al 1999).…”

Section: Binding Of Sapp To Cell Surfacesmentioning

confidence: 95%

The biological role of the Alzheimer amyloid precursor protein in epithelial cells

Schmitz

Tikkanen

Kirfel

et al. 2001

Histochem Cell Biol

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Proteolytic processing of the Alzheimer amyloid precursor protein (APP) results in the generation of at least two distinct classes of biologically relevant peptides: (1) the amyloid beta peptides which are believed to be involved in the pathogenesis of Alzheimer's disease and (2) the soluble N-terminal ectodomain (sAPP) which exhibits a protective but as yet ill-defined effect on neurons and epithelial cells. In this report we present an overview on the functions of sAPP as an epithelial growth factor. This function involves specific binding of sAPP to membrane rafts and results in signal transduction and various physiological effects in epithelial cells as different as keratinocytes and thyrocytes. At nanomolar concentrations sAPP induces a two to fourfold increase in the rate of cell proliferation and cell migration. Specific inhibition of APP expression by antisense techniques results in decreased sAPP release and in reduced proliferative and motogenic activities. Proliferation and migration are known to be part of complex processes such as wound healing which, therefore, might be facilitated by the growth factor function of sAPP.

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Section: Binding Of Sapp To Cell Surfacesmentioning

confidence: 95%

The biological role of the Alzheimer amyloid precursor protein in epithelial cells

Schmitz

Tikkanen

Kirfel

et al. 2001

Histochem Cell Biol

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“…Among the many available protein crosslinking agents, glutaraldehyde has undoubtedly found the widest application in various fields such as histochemistry (13), microscopy (1,4,5), cytochemistry (6), leather tanning industry (7,8), enzyme technology (913), chemical sterilization (14), and biomedical (15) and pharmaceutical sciences (16). Glutaraldehyde, a linear, 5-carbon dialdehyde, is a clear, colorless to pale straw-colored, pungent oily liquid that is soluble in all proportions in water and alcohol, as well as in organic solvents.…”

Section: Introductionmentioning

confidence: 99%

Glutaraldehyde: Behavior in Aqueous Solution, Reaction with Proteins, and Application to Enzyme Crosslinking

et al. 2004

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Glutaraldehyde possesses unique characteristics that render it one of the most effective protein crosslinking reagents. It can be present in at least 13 different forms depending on solution conditions such as pH, concentration, temperature, etc. Substantial literature is found concerning the use of glutaraldehyde for protein immobilization, yet there is no agreement about the main reactive species that participates in the crosslinking process because monomeric and polymeric forms are in equilibrium. Glutaraldehyde may react with proteins by several means such as aldol condensation or Michael-type addition, and we show here 8 different reactions for various aqueous forms of this reagent. As a result of these discrepancies and the unique characteristics of each enzyme, crosslinking procedures using glutaraldehyde are largely developed through empirical observation. The choice of the enzyme-glutaraldehyde ratio, as well as their final concentration, is critical because insolubilization of the enzyme must result in minimal distortion of its structure in order to retain catalytic activity. The purpose of this paper is to give an overview of glutaraldehyde as a crosslinking reagent by describing its structure and chemical properties in aqueous solution in an attempt to explain its high reactivity toward proteins, particularly as applied to the production of insoluble enzymes.

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“…Glutaraldehyde may react with amino groups in different ways, depending on temperature, protein concentration in the tissue, pH, concentration of the glutaraldehyde and the ionic strength of the fixative (Hopwood, 1972). The rate of the crosslinking reaction is also governed by the above variables.…”

Section: Introductionmentioning

confidence: 99%

Penetration rate of glutaraldehyde in various buffers into plant tissue and gelatin gels

Coetzee

Merwe

1985

Journal of Microscopy

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SUMMARY The choice of buffer vehicle during fixation with glutaraldehyde influences not only the rate of crosslinking in a bovine serum albumin model system, but also the penetration rate of the glutaraldehyde into plant tissue and gelatin gels. The rate of penetration into plant tissue and the crosslinking rate of bovine serum albumin by glutaraldehyde are independent of each other in any one buffer. Buffer penetration into gelatin gels is not as fast as glutaraldehyde penetration into the same gels. The highest rate of crosslinking of bovine serum albumin by glutaraldehyde is obtained with a buffer vehicle consisting of Na2HPO4 and NaH2PO4. Glutarldehyde concentrations of 2–3% are advised for routine fixation of plant material.

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Theoretical and practical aspects of glutaraldehyde fixation

Cited by 213 publications

References 115 publications

The biological role of the Alzheimer amyloid precursor protein in epithelial cells

The biological role of the Alzheimer amyloid precursor protein in epithelial cells

Glutaraldehyde: Behavior in Aqueous Solution, Reaction with Proteins, and Application to Enzyme Crosslinking

Penetration rate of glutaraldehyde in various buffers into plant tissue and gelatin gels

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