摘要:

AbstractThe degree of interaction between protein and stains depends in large part upon the protein's content of basic amino acids. We have compared “staining” characteristics of Coomassie Brilliant Blue R‐250 with a dye that was rendered radioactive by neutron bombardment. The protein‐dye intensities were compared to amino acid composition as in our previous study (Electrophoresis1986,7, 327–332). While the data for “colored” dye reflected involvement of basic amino acid residues, quite unexpectedly, the radioactive dye appeared to act in part with the acidic amino acids. This may be due to enhanced binding of dye impurities or radiol

ISSN:0173-0835    

DOI:10.1002/elps.1150081202

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

年代:1987

数据来源: WILEY

摘要:

AbstractA rapid method for determining the reactivity of murine monoclonal antibodies with apolipoprotein A‐I (Apo A‐I) isoforms has been developed. The method consists of a rapid (1–2 h) prescreening of fusion well culture fluids by using an automated immunofluorescent assay (IF A) to identify fusion well cultures producing monoclonal antibody against Apo A‐I and high‐density lipoprotein (HDL). Immediately following the IFA procedure, immunodetection of the reactivity pattern of selected fusion well culture fluids is determined by using transblotted isoforms of Apo A‐I previously separated by isoelectric focusing. Strips are prepared and stored at 4 °C in protein containing blocking solution before use. Preselected culture fluids could be tested within 6 h for qualitative reactivity with Apo A‐I isoforms. Using this procedure we selected several monoclonal antibody‐producing cultures for further studies on HDL metabolism based on their differing reactivities with Apo A‐I isoforms. This technique should be easily transferable to other laboratories for assessing proteins having multiple isoforms since the crucial steps of separation and blotting can be optimized and performed before actual testing for a

ISSN:0173-0835    

DOI:10.1002/elps.1150081203

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

年代:1987

数据来源: WILEY

摘要:

AbstractThe effect of sialic acid on the electrophoretic mobility (surface charge) of mouse splenic lymphocytes was investigated. On treatment of whole spleen and T cells with neuraminidase, their electrophoretic mobilities decreased to one‐half of the initial values. Calculation of the surface charge from the electrophoretic mobility of lymphocytes before and after treatment with the enzyme indicated that one‐half of the negative surface charge, which determines the electrophoretic mobility, was due to neuraminidase‐sensitive sialic acid in mouse lymphocytes. However, the calculated decrease in the surface charge did not coincide with the amount of sialic acid released from the cell membrane on the enzyme treatment of lympho

ISSN:0173-0835    

DOI:10.1002/elps.1150081204

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

年代:1987

数据来源: WILEY

摘要:

AbstractA novel and unique protein pattern for alpha‐2‐HS‐glycoprotein (A2HS) was found in the sera from two unrelated Afro‐Caribbean donors after isoelectric focusing followed by immunofixation. In the native state the variant bands were located between A2HS 7 and A2HS 10 bands. The desialized samples showed that a cathodal variant band was indistinguishable from the A2HS 5 band. These findings indicated that both these donors possessed a variant phenotype of A2HS determined by a new alleleA2HS*11, in combination with the commo

ISSN:0173-0835    

DOI:10.1002/elps.1150081205

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

年代:1987

数据来源: WILEY

摘要:

AbstractThree two‐dimensional electrophoretic protein patterns (fluorograms) showing a representative spectrum of different qualities of resolution (spots well separated, spots embedded in “streaks” and spots of low contrast to the background) were selected to ascertain the optimum of the parameters of a computer program used for spot detection with a video camera/computer system. A “visual norm‐pattern” was constructed on the basis of the results obtained by visual evaluation of the three patterns by several test persons. The visual norm‐pattern (a graphic spot pattern) consisted of two classes of spots: “visible spots” (spots detected by all test persons) and “invisible but true spots” (spots detected by all persons, but only if the exposure time of the X‐ray film was increased). This pattern was used as a standard pattern for automatic evaluation. The automatic evaluation of the three test patterns was performed under various conditions: In the computer program two parameters critical for spot detection, parameterP1(responsible for the separation of the spots) andP2(responsible for the size of the spots), were varied over a broad range. In this way a third class of spots, “invisible, artifactual spots”, was revealed. The three spot classes could not be distinguished by means of the automatic system. Therefore, the visual norm‐pattern was used to adjust parametersP1andP2at an optimum level of sensitivity. The optimum values found forP1andP2allowed us to detect all the visible spots of a two‐dimensional protein pattern, and, in addition, a portion of the invisible but true spots, while none of the artifactual spots was included in the computer‐produced patterns. TheP‐values chosen could be used for two‐dimensional protein patterns of different qualities. Slight deviations of theP‐

ISSN:0173-0835    

DOI:10.1002/elps.1150081206

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

年代:1987

数据来源: WILEY

摘要:

AbstractBovine fetal and adult serum/plasma proteins were analyzed by two‐dimensional electrophoresis in the absence of denaturants. Bovine serum proteins were identified by electrophoretic transfer from gel slabs followed by immunochemical staining. Amine oxidase was identified by activity staining. Antisera against human plasma proteins were employed for the immunochemical identification, together with antisera to bovine proteins. Developmental changes of fetal serum proteins including α‐fetoprotein were visual

ISSN:0173-0835    

DOI:10.1002/elps.1150081207

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

年代:1987

数据来源: WILEY

摘要:

AbstractMid‐morning specimens of unstimulated whole saliva were collected from 160 apparently healthy individuals and analyzed by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis following sample preparation with or without 2‐mercaptoethanol. Coomassie Brilliant Blue R‐250 staining revealed blue and pink‐violet salivary protein components. The latter, detected in approximately 60 % of the individuals, formed a limited range of patterns combining 2–8 of 11 diffuse zones ofMr10 000–100 000. The pink‐violet components were unaffected by 2‐mercaptoethanol, urea or prior centrifugation of the sample and were not invariably present or absent in repeat samples from the same individual. The patterns of the blue staining components were more reproducible for the purpose of human individualisation but were affected by 2‐mercaptoethanol or prior centrifugation of the sample. The patterns of both the blue and pink‐violet components were not affected by sample storage at −70 °C. Only 5 μL of saliva were required for analysis giving excellent resolution over

ISSN:0173-0835    

DOI:10.1002/elps.1150081208

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

年代:1987

数据来源: WILEY

摘要:

AbstractA procedure is described to improve the properties of dried gels, containing immobilized pH gradients, for rehydration in the presence of urea. Triton X‐100, reducing agents and carrier ampholytes. When gels are dried by a fan in ambient atmosphere for the shortest time interval required to achieve an evenly flat surface, rehydration in the presence of 8 mol/L urea and 30 or 5 mL/L of Triton X‐100 is completed within 4 or 1 h, respectively. These rehydration properties can be maintained by storing the dried and sealed gels at −

ISSN:0173-0835    

DOI:10.1002/elps.1150081209

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

年代:1987

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150081210

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

年代:1987

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150081201

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

年代:1987

数据来源: WILEY