Studies on Cystic Fibrosis Using Isoelectric Focusing. I. An Assay for Detection of Cystic Fibrosis Homozygotes and Heterozygote Carriers from Serum

Wilson, Gregory B.; Fudenberg, H. Hugh; Jahn, Theodore L.

doi:10.1203/00006450-197508000-00005

Cited by 58 publications

(38 citation statements)

References 9 publications

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“…It is of interest to note that the same methods of sample collection and storage that we use (8) also result in C F homozygote and heterozygote carrier samples which are positive for C F ciliary dyskinesia factor (CDF) (3) as detected by a modified rabbit tracheal bioassay (10, I I), whereas normal healthy control sera are CDF negative. ), it is obvious that neither group has reproduced the protein banding patterns presented in either of our previous reports (8,9). The following points are evident upon comparing their figure ( 5 ) to those previously published (8).…”

supporting

confidence: 61%

“…), it is obvious that neither group has reproduced the protein banding patterns presented in either of our previous reports (8,9). The following points are evident upon comparing their figure ( 5 ) to those previously published (8). (1) The pH gradients are not the same, (2) the location of the sample pads relative to the anode is not the same and, (3) the protein banding patterns are totally different.…”

supporting

confidence: 49%

“…These investigators were able to demonstrate a small molecular weight protein unique to sera from C F homozygotes and heterozygotes by performing a second step electrophoresis utilizing the serum protein fraction which focuses between pH 7 and 9. Curiously enough neither Smith et al (4) nor Thomas et al (5) could reproduce either Altland's findings (1) or ours (8,9).…”

mentioning

confidence: 72%

“…In the letter by Smith et al (4) and an abstract of work presented by Thomas et al (5), two attempts at reproducing our assay are described. Both groups of investigators claim to have analyzed C F homozygote and normal control sera using electrofocusing methods exactly analogous to those published (8,9) with the obvious exception that neither group used the electrofocusing apparatus used in our method (2,6,8,9). Smith et a/.…”

mentioning

confidence: 99%

“…We have previously described an assay for the detection of individuals homozygous or heterozygous for cystic fibrosis (CF), which employs analytical electrofocusing in thin layer polyacrylamide gels to examine sera for a cystic fibrosis protein (CFP) and other protein markers (bands B, C, D) that we feel are diagnostic for C F (8,9). In the letter by Smith et al (4) and an abstract of work presented by Thomas et al (5), two attempts at reproducing our assay are described.…”

mentioning

confidence: 99%

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Letter to the Editor

Arnaúd

et al. 1976

Pediatr Res

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supporting

confidence: 61%

supporting

confidence: 49%

mentioning

confidence: 72%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Letter to the Editor

Arnaúd

et al. 1976

Pediatr Res

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Cystic fibrosis proteins detected by electrophoretic techniques

Guy-Crotte¹,

Barthe²,

Figarella³

1989

Electrophoresis

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For the detection of the cystic fibrosis protein (CFP) in serum of cystic fibrosis (CF) carriers, thin-layer polyacrylamide gel isoelectric focusing proved inappropriate as a diagnostic test, but was useful for screening fractions on purification of CFP by chromatofocusing on a Mono P column. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis an Mr 12,000 protein (P12) was found in most CFP-positive sera, indicating good correlation between these two CF-associated proteins. Detection of the P12 protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was well reproducible and less delicate than IEF. The technique was also used to purify P12 from serum by two successive preparative electrophoresis steps in a 7.5-15% gradient and 15% homogeneous gel. The use of silver staining revealed that P12, which was present in all sera of CF patients and carriers with variable intensities, was also present in trace amounts in normal sera.

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Detection of cystic fibrosis protein by isoelectric focusing and silver staining in serum, dried whole blood, and cultured fibroblasts

Vladutiu

Ryan

Cropp

1988

Clinical Laboratory Analysis

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Experimental conditions were developed for visualizing cystic fibrosis protein (CFP) in cultured fibroblasts from patients with cystic fibrosis (CF) while improving the visualization of CFP in serum. Serum, whole dried blood and cultured skin fibroblast extracts derived from CF homozygotes, heterozygotes, and normal individuals were submitted to isoelectric focusing (IEF) in polyacrylamide gels using a pH gradient of 7.1 to 9.3 and stained for protein with silver. The results showed an improvement in the separation of bands in the cathodal region over that obtained with a pH gradient of 3.5 to 10 and a better visualization of CFP bands than provided with Coomassie blue. Serum IgG could be removed from the cathodal region by including Protein A on the gel during IEF, thus enabling better visualization of CFP bands. With cultured fibroblast extracts, CFP was only observed when gels were stained with silver. This study presents improved sensitivity of a potential diagnostic tool in CF and its use with cultured skin fibroblasts.

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Studies on Cystic Fibrosis Using Isoelectric Focusing. I. An Assay for Detection of Cystic Fibrosis Homozygotes and Heterozygote Carriers from Serum

Cited by 58 publications

References 9 publications

Letter to the Editor

Letter to the Editor

Cystic fibrosis proteins detected by electrophoretic techniques

Detection of cystic fibrosis protein by isoelectric focusing and silver staining in serum, dried whole blood, and cultured fibroblasts

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