摘要:

AbstractWe describe a modification in the sample application mode for isoelectric focusing with immobilized pH gradients. Instead of being applied at the surface of the gel in a sample cup, the sample is introduced into the gel during the immobilized pH gradient strip rehydration step. This modification implies the use of low percentage gels (below 3.5%T) and specially designed, but simple, rehydration chambers. The main advantages are a uniform resolution without side effects and the possibility of handling large sample volumes (500 μL for a standard 3 × 160 × 0.5 mm strip), allowing micropreparative work (milligram samples) with a simple experimental desi

ISSN:0173-0835    

DOI:10.1002/elps.11501501223

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractIn an attempt to generalize previous observations (Jaenickeet al., Appl. Environ. Microbiol. 1988,54, 2375–2380) and to find a convenient model system for studies of the pressure response, we tested the suitability ofEscherichia coliandThermotoga maritima(bacteria), and of five different eukaryotic species including the filamentous fungiAsteromyces cruciatusandDendryphiella salina., and the marine yeastsDebaryomyces hansenii, Rhodosporidium sphaerocarpum, andRhodotorula rubra.Using two‐dimensional polyacrylamide gel electrophoresis, detailed investigations on the pressure response were carried out withE. coliandRhodosporidium sphaerocarpum. In the former organism, major pressure response proteins could not be detected, although there are significant differences in expression of some proteins as well as some minor components that are found in all of the high pressure cell extracts but not in extracts from cultures grown at atmospheric pressure. InRhodosporidium sphaerocarpum, no change in protein expression patterns was observed between 0.1 and 20 MPa. However, approaching the limit of viability of 50 MPa, additional protein spots became detectable at 45 MPa. This finding correlates with the observation of abnormal growth forms of the organism at this pressure (Lorenz, R.et al.manuscript in preparati

ISSN:0173-0835    

DOI:10.1002/elps.11501501224

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractWe have utilized two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE) coupled with silver stain to identify cellular proteins in human non‐Hodgkin's B‐lymphoma (NHL). Five cell lines (SKDHL2B, WSU‐DLCL2, WSU‐NHL, WSU‐FSCCL and SKLN1), representing four different NHL maturational stages and a normal Epstein‐Barr virus (EBV)‐transformed line of B‐cell origin (SKLN1) were studied. The NHL lines were immunophenotyped using flow cytometry with lineage associated monoclonal antibodies. Whole cell lysates of the cell lines were subjected to 2‐D PAGE analyses. The gels were analyzed with an image scanning computer and the qualitative differences of protein patterns were studied. Results revealed great similarities in patterns of the NHL lines. A master map containing common NHL‐protein spots was constructed. When the map of each tumor line was compared to the master map, several protein spots were associated with each NHL‐grade. Search for these proteins in the normal EBV‐transformed B‐cell line showed that only one of the proteins (S3;Mr/pI19/5.9) was present. Proteins that were detected in malignant NHL, but not in the normal EBV‐line, could provide important information regarding the human NHL B‐lymphocyte databases. Whether or not these proteins are definite malignant markers to distinguish between different NHL maturational stages needs further exploration through el

ISSN:0173-0835    

DOI:10.1002/elps.11501501225

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractRelative abundar ce and solubility of proteins from etiolated coleoptiles of maize were investigated using two‐dimensional electrophoresis (2‐D PAGE). Automatic quantification of silver‐stained polypeptides on replicate 2‐D gels made it possible to test the linearity of the relationship between spot integrated optical density and protein amount in the range from 15 μg to 135 μg per gel, in two inbred lines. A linear response was found for more than 60% of the spots in each genotype. When a linear response was found in both lines for a given spot, the slope values were similar in 94% of the cases, indicating the reliability of silver staining for polypeptide quantification. The parameters of the curves allowed the definition of protein classes of different abundances that could be compared for genetic variability between the two lines. From a comparison between standard 2‐D patterns (trichloroacetic acid‐acetone extracted proteins) and patterns obtained from Tris buffer extracted proteins, it appeared that 92% of the proteins visualized in the standard gels were soluble. No difference in genetic variability, ether qualitative or quantitative, was evidenced between the various classes of abundance, or between soluble and insol

ISSN:0173-0835    

DOI:10.1002/elps.11501501226

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.11501501227

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150150101

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY