Protein singly-labeled

The respective marker proteins, single or mixed, are dissolved in Soln. A to a concentration of 20 mg/ml at RT. Then 0.5-1 vol. of Soln. B are added and the mixture is heated to 60 °C for 5 min. After cooling, the labeled proteins are lyophilized without further purification and dissolved again in Soln. C to a concentration of 20 mg/ml. If the assay was larger than 0.1 ml, the product is aliquoted and stored at -20 °C. 

Morphological approaches in biomedical research can include a wide range of microscopes, but today typically employ immunocytochemistry that can give us information about individual liver cells containing the specific enzyme. Immunocytochemistry uses antibodies to bind proteins and labels to show protein s location. If, for example, the enzyme is a marker for inflammation, then the location of cells with this enzyme tells us which cell types have the inflammatory response. Thus, immunocytochemistry methods are broadly defined as individual studies of single cells or cell groups. The resulting data tell us about location of the enzyme. 

Fig. 4 Cartoon representation of EPR distance measurements, (a) Doubly labeled monomeric proteins give rise to intra- and intermolecular spin-spin interactions. In order to determine intramolecular distances, experimental data has to be corrected for intermolecular contributions, (b) Intermolecular distance measurements using singly labeled proteins in protein oligomers or aggregates. Multiples of the distance are also expected. This may be even more complicated for different types of aggregation and can be analyzed by studying a series of samples with increasing content of non-labeled molecules (diamagnetic dilution), (c) To measure intramolecular distances within oligomers/aggregates, a mixture of doubly labeled and non-labeled proteins can be used...

The non-interacting powder pattern which is required for distance analysis is experimentally accessible by measuring EPR spectra of samples containing either singly labeled proteins or, in the case of intermolecular distances, containing both labeled and unlabeled proteins ( diamagnetic dilution ) to avoid interspin distances below 2.0 nm. 

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