A novel and non-destructive approach for ZooMS analysis: ammonium bicarbonate buffer extraction

2011 ◽  
Vol 3 (3) ◽  
pp. 281-289 ◽  
Author(s):  
Nienke Laura van Doorn ◽  
Hege Hollund ◽  
Matthew J. Collins
Author(s):  
Douglas R. Keene ◽  
B. Kerry Maddox ◽  
Marie B. Spurgin ◽  
Lynn Y. Sakai ◽  
Robert W. Glanville

A mouse monoclonal antibody was used to identify beaded aggregates found in guanidine extracts of human amnion as assemblies of fibrillin molecules. These aggregates were also shown to be a major component of extracellular matrix microfibrils. We further demonstrated that the periodicity of these aggregates can be increased when subjected to mechanical stress.Human amnion was extracted with guanidine and the extracted material purified using ion exchange and molecular sieve chromatography. A high molecular weight fraction was precipitated by dialyzing against dilute acetic acid. Part of the precipitate was suspended in 0.2 M ammonium bicarbonate buffer and rotary shadowed. A second portion was resuspended in culture medium containing antibody which recognizes matrix microfibrils, diluted 1:5 in ammonium bicarbonate and reacted for 120 minutes at room temperature. Antibody labeled precipitate was washed by repeated pelleting and resuspension in buffer and then incubated in Janssen GAM 5 nm gold conjugate for 60 minutes at room temperature.


Molecules ◽  
2020 ◽  
Vol 25 (24) ◽  
pp. 5982
Author(s):  
Sorel Tchewonpi Sagu ◽  
Eva Landgräber ◽  
Michal Rackiewicz ◽  
Gerd Huschek ◽  
Harshadrai Rawel

Sorghum is of growing interest and considered as a safe food for wheat related disorders. Besides the gluten, α-amylase/trypsin-inhibitors (ATIs) have been identified as probable candidates for these disorders. Several studies focused on wheat-ATIs although there is still a lack of data referring to the relative abundance of sorghum-ATIs. The objective of this work was therefore to contribute to the characterization of sorghum ATI profiles by targeted proteomics tools. Fifteen sorghum cultivars from different regions were investigated with raw proteins ranging from 7.9 to 17.0 g/100 g. Ammonium bicarbonate buffer in combination with urea was applied for protein extraction, with concentration from 0.588 ± 0.047 to 4.140 ± 0.066 mg/mL. Corresponding electrophoresis data showed different protein profiles. UniProtKB data base research reveals two sorghum ATIs, P81367 and P81368; both reviewed and a targeted LC–MS/MS method was developed to analyze these. Quantifier peptides ELAAVPSR (P81367) and TYMVR (P81368) were identified and retained as biomarkers for relative quantification. Different reducing and alkylating agents were assessed and combination of tris (2 carboxyethyl) phosphine/iodoacetamide gave the best response. Linearity was demonstrated for the quantifier peptides with standard recovery between 92.2 and 107.6%. Nine sorghum cultivars presented up to 60 times lower ATI contents as compared to wheat samples. This data suggests that sorghum can effectively be considered as a good alternative to wheat.


Author(s):  
J W Steeds

There is a wide range of experimental results related to dislocations in diamond, group IV, II-VI, III-V semiconducting compounds, but few of these come from isolated, well-characterized individual dislocations. We are here concerned with only those results obtained in a transmission electron microscope so that the dislocations responsible were individually imaged. The luminescence properties of the dislocations were studied by cathodoluminescence performed at low temperatures (~30K) achieved by liquid helium cooling. Both spectra and monochromatic cathodoluminescence images have been obtained, in some cases as a function of temperature.There are two aspects of this work. One is mainly of technological significance. By understanding the luminescence properties of dislocations in epitaxial structures, future non-destructive evaluation will be enhanced. The second aim is to arrive at a good detailed understanding of the basic physics associated with carrier recombination near dislocations as revealed by local luminescence properties.


Author(s):  
R.F. Sognnaes

Sufficient experience has been gained during the past five years to suggest an extended application of microreplication and scanning electron microscopy to problems of forensic science. The author's research was originally initiated with a view to develop a non-destructive method for identification of materials that went into objects of art, notably ivory and ivories. This was followed by a very specific application to the identification and duplication of the kinds of materials from animal teeth and tusks which two centuries ago went into the fabrication of the ivory dentures of George Washington. Subsequently it became apparent that a similar method of microreplication and SEM examination offered promise for a whole series of problems pertinent to art, technology and science. Furthermore, what began primarily as an application to solid substances has turned out to be similarly applicable to soft tissue surfaces such as mucous membranes and skin, even in cases of acute, chronic and precancerous epithelial surface changes, and to post-mortem identification of specific structures pertinent to forensic science.


2013 ◽  
Vol 64 (2) ◽  
pp. 21001 ◽  
Author(s):  
Jean-Luc Bodnar ◽  
Jean-Jacques Metayer ◽  
Kamel Mouhoubi ◽  
Vincent Detalle

1984 ◽  
Vol 72 (8-9) ◽  
pp. 339-343
Author(s):  
René Hoeg ◽  
Lars Taarnskov
Keyword(s):  

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