All issues
Successor
Volume 42, Issue 3
Displaying 1-9 of 9 articles from this issue
- |<
- <
- 1
- >
- >|
-
Jun-ichi Asakawa1998 Volume 42 Issue 3 Pages 145-154
Published: August 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSA vertical giant gel two-dimensional electrophoresis (2-DE) system has been developed and applied to the restriction landmark genomic scanning (RLGS; 2 DE of end labeled genomic DNA fragments). A unique gel holder for the first-dimension agarose disc gel was made from teflon-tubing and glass tube. The development of the disc gel system resulted in high-resolution separation of DNA fragments with enhanced reproducibility. A new device for the in-gel restriction enzyme digestion of the fractionated DNA fragments was also made from teflon-tubing. After optimization of the sample preparation and electrophoresis conditions, the gel quality reached a level such that the electrophoresis patterns derived from a single DNA sample gave distribution patterns of spots able to be superimposed. A protocol and the equipment for the 2-DE are described.View full abstractDownload PDF (5054K) -
Youji Shimazaki, Takashi Manabe1998 Volume 42 Issue 3 Pages 155-159
Published: August 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSUsing microscale multisample two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) in the absence of denaturing agents, eight protein samples can be simultaneously analyzed and compared. In the present study, this method was applied for the analysis of chicken serum, chicken egg yolk and chicken egg albumen proteins, and the 2-D PAGE pattern of chicken yolk and albumen were compared with that of chicken serum. The spot locations (pI and Mr) of yolk proteins resembled with that of chicken serum, but those of albumen proteins were considerably different from those of chicken serum. Employing the techniques of electrophoretic blotting and immunochemical staining, immunoglobulin G (IgG), third component of complement (C3) and transferrin were identified in the patterns of chicken serum and yolk proteins.View full abstractDownload PDF (3192K) -
Tadashi Hoshino, Makoto Matsushita, Tsutomu Irino, Kazunari Kumasaka, ...1998 Volume 42 Issue 3 Pages 161-167
Published: August 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSAt least 4 isoforms of intestinal alkaline phosphatase (ALP) have been identified in human, which are common intestinal ALP, intestinal ALP variant, blood group-dependent high-molecular intestinal ALP, and fetal intestinal ALP. These isoforms can be clearly separated by polyacrylamide gel electrophoresis (PAGE). However, comparative studies on electrophoretic mobilities of these isoforms which are separated by cellulose acetate membrane and agarose gel have not been done yet. On the cellulose acetate membrane electrophoresis, a relatively tightened band of intestinal ALP variant may be presented at the position of liver/bone ALP (α2-α2β position) and a broad band of the blood group-dependent high-molecular intestinal ALP may be exhibited at the position overlap from bone ALP to common intestinal ALP (α2β-β position). On the agarose gel electrophoresis, a tightened band of the intestinal ALP variant and/or the blood group-dependent high-molecular intestinal ALP was possible to detected around bone ALP (α2β position).View full abstractDownload PDF (3502K) -
Mayumi Tsumura, Yoshito Ueno, Kazuaki Morio, Shoji Kuwata, Takashi Kin ...1998 Volume 42 Issue 3 Pages 169-174
Published: August 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSWe found frequently an atypical alkaline phosphatase (ALP) band which detected at the upper region of the separating gel (atypical ALP-1) by the method of polyacrylamide disc gel (PAG) electrophoresis in the patients with certain renal failure. Although most of clinicians have demonstrated the assay of β-N-acetylglucosaminidase (NAG) and β2-microglobulin (β2MG) as the useful marker for advanced renal failure, we then examined the relationship between the levels of β2MG and the atypical ALP-1 in the patients with chronic renal failures. In particular, we studied the relationship between atypical ALP and β2MG in two groups of chronic renal failure, one is with diabetes mellitus (DM) and the other is without diabetes mellitus (non DM). From the present results, higher appearances of the atypical ALP-1 in the serum or urine of the patient with DM nephropathy groups were well accorded to the levels of serum or urinary β2MG in the matched patient group. Taken together, the blood or urinary appearance of atypical ALP-1 and the level of β2MG may be a good tool for the pathological conditions in the patients with diabetic nephropathy.View full abstractDownload PDF (1900K) -
Yoko Toyoda, Yoshito Ueno, Narumi Takashina, Mayumi Tsumura, Shoji Kuw ...1998 Volume 42 Issue 3 Pages 175-179
Published: August 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSWe detected the extra band cathodal to the electrophoretic origin through the analysis of CK isozyme of 346 healthy subjects. This band was observed in all the fresh sera stored at 4°C less than 24h. Biochemical analysis disclosed that neither adenylate kinase nor immunoglobulin-bound CK generated this band. The shift of the extracts migration after the incubation with urea implied that this extra band was derived from mitochondrial CK. We furthermore confirmed the high activation energy of this band (111.75kJ/mol) which was compatible with that of mitochondrial CK. These results strongly support that this extra band was identical with mitochondrial CK. In conclusion presumed mitochondrial CK was observed in the fresh sera of all the healthy subjects. Ordinary cell turnover may be postulated to contribute to this extra band but further study will be needed to define the precise features of this band.View full abstractDownload PDF (3021K) -
Shigemi Hashimoto, Motofumi Hiyoshi, Takayuki Takubo, Shinichi Tagawa, ...1998 Volume 42 Issue 3 Pages 181-184
Published: August 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSAn Amplicor PCR series (Roche, Branchburg, NJ, USA) was developed to make the PCR technology available in hospital as a routine test, and is prevailing for the diagnosis of Mycobacterium tuberculosis, Cytomegalovirus (CMV), etc. In these circumstances, we sometimes experience a discrepancy between the result of Amplicor diagnosis and the patients' clinical findings. A method is described here to recheck the Amplicor results by performing agarose gel electrophoresis with the PCR products obtained from Amplicor. Amplicor CMV PCR test was used for this study. When the PCR products were electrophoresed without any purification, bands of amplified CMV DNA were not electrophoresed precisely. On the other hand, when the products were purified by the Qiagen blood kit (Hiden, Germany), the PCR products were electrophoresed properly. We concluded that the best way to recheck the Amplicor results was to run electrophoresis after purification of PCR products.View full abstractDownload PDF (1771K) -
Yuriko Kurihara, Kazunori Yokoyama, Kiyoko Shiba, Sachiko Kamei1998 Volume 42 Issue 3 Pages 185-188
Published: August 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSWe analyzed glycosaminoglycans and proteins in the exudate of an esthesioneuroblastoma. The total protein concentration was 44.0g/l. Exudate concentrations of IgG, IgA, and IgM were 6.0g/l, 2.6g/l, and 12.6g/l, respectively. IgM λ-type M-protein was found in the exudate sample. The glycosaminoglycan contents were 32.4% hyaluronic acid and 39.3% heparan sulfate. Both existed in high concentrations in the exudate.View full abstractDownload PDF (3219K) -
Motowo Nakajima, Minako Toyoshima, Sadako Yamagata, Tatsuya Yamagata1998 Volume 42 Issue 3 Pages 189-196
Published: August 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSCancer is the major mortality in most advance countries and the development of efficient diagnosis and prognosis tools for cancer therapy is one of the major goals of cancer research. Highly sensitive, reliable and convenient assays using non-invasive means are required to invent new cancer diagnostic protocols. Zymography of serum and plasma samples by electrophoresis and incubation in a substrate-embedded polyacrylamide gel is suitable for such routine diagnostic assays. Zymography is also a unique method to discover unidentified glycosidases capable of specifically cleaving a certain type of glycosaminoglycans such as hyaluronic acid and chondroitin sulfate. Here we first report the new application of zymography to detect novel glycosaminoglycanases in human malignant cancers using chemically-modified chondroitin sulfate embedded in polyacrylamide gels. Secondly, we demonstrate a correlation between plasma matrix metalloproteinase activity and tumor progression with metastasis using gelatin zymography and then discuss a possible application of zymography to cancer diagnosis.View full abstractDownload PDF (6925K) -
Kiyohito Shimura1998 Volume 42 Issue 3 Pages 197-202
Published: August 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSThe use of polyelectrolytes as mobile affinity matrices in electrophoresis has led to the development of a specific separation method for biological molecules, affinophoresis. The conjugate of a polyelectrolyte and an affinity ligand is called an affinophore. Electrophoresis of proteins in the presence of an affinophore results in a change in the mobility of a specific protein by the formation of a specific complex through the affinity of the protein to the ligand. Polylysine is useful as a base polymer of affinophores and has been used successfully as an anionic matrix after succinylation. Affinophoresis of proteases, lectins and antibodies has been carried out in agarose gel and the mobility of the proteins having affinity to each ligand was specifically changed. Two-dimensional affinophoresis, in which an affinophore was included only in the second-dimensional electrophoresis, was effective for the identification of the molecule having affinity to the affinophore in a complex mixture. The use of capillary electrophoresis highlighted the relevance of affinophoresis in quantitative analyses of molecular interactions, allowing the determination of affinity constants with small amount of samples and with high precision.View full abstractDownload PDF (3634K)
- |<
- <
- 1
- >
- >|