• Archives of Pharmacal Research
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• v.25 no.5
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• pp.704-708
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• 2002

A fast and sensitive protein staining method in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using both an acidic dye, Coomassie Brilliant Blue R-250 (CBBR) and a basic dye, Neutral Red (NR) is described. It is based on a counter ion-dye staining technique that employs oppositely charged two dyes to form an ion-pair complex. The selective binding of the free dye molecules to proteins in an acidic solution enhances the staining effect of CBBR on protein bands, and also reduces gel background. It is a rapid staining procedure, involving fixing and staining steps with short destaining that are completed in about 1 h. As the result, it showed two to fourfold increase in sensitivity comparing with CBBR staining. The stained protein bands can be visualized at the same time of staining.

• Bulletin of the Korean Chemical Society
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• v.23 no.11
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• pp.1511-1512
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• 2002

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.9
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• pp.57-63
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• 2020

To develop simple acrosome staining of horse spermatozoa, this study tested the binding properties of Coomassie brilliant blue G or R on the sperm smears after 3.7% paraformaldehyde (PF) or 35% methanol (MT) fixation. After being fixed with PF and stained with 0.05, 0.1, or 0.2 % of CBB G or R for 2 min, horse spermatozoa were examined for their intact acrosome status. The intact acrosome of fresh horse spermatozoa were 62.6% and 61.5% with 0.05% of the G and R CBB solution, but 80.2 and 79.7% with G type and 78.1 and 76.0% with R type. On the other hand, when MT was used for fixation, the acrosome reacting sperm ratio was 3.5%, but was 9.0% in the case of PF. These results show that the intact acrosome of horse sperm could be judged using a 0.1~0.2% CBB G or R staining technique. PF would be an essential fixative for examining acrosome reacting horse spermatozoa. This method could be used to identify sperm with a damaged acrosome during low-temperature storage or cryopreservation for artificial insemination of horses.

• Proceedings of the PSK Conference
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• 2000.10a
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• pp.194.2-195
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• 2000

• BMB Reports
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• v.34 no.6
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• pp.531-536
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• 2001

Three zymographic techniques using casein, fibrin, and gelatin as substrates in SDS-PAGE were compared based on three aspects: (1) The proteolytic pattern of extracellular enzymes from the three bacterial strains, Bacillus sp. DJ-1, DJ-2, and DJ-3. (2) The enzymatic sensitivity of their activity on zymogram gels. (3) The stability of stained zymogram gels with Coomassie brilliant blue in the destaining solution. There was no significant difference on the pattern of extracellular enzymes from the three strains. The bands in the fibrin gel were clearer and more distinct from the extensive destaining process. It was also shown that the gelatin gel revealed the highest enzymatic sensitivity among the three gels, based on the densitometric analysis. In the casein gel, a trace that could be mistaken as a proteolytic band appeared around 40-50 kDa.

• Molecular & Cellular Toxicology
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• v.4 no.3
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• pp.253-259
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• 2008

Human follicular fluid (HFF) is the in vivo microenvironment for oocyte maturation and includes a variety of proteins that could be involved in oocyte development and fertilization. We therefore used a proteomic approach to identify new HFF proteins. HFF from mature human follicles was obtained from five women following oocyte collection for in vitro fertilization (IVF). Ethanol-precipitated HFF run on two-dimensional gel electrophoresis (2DE) produced approximately 250 Coomassie brilliant blue-stained spots, 64 of which were identified using matrix-assisted laser desorption/ionization-mass spectrometry (MALDIMS). In this study, several proteins including complement factor H, inter-${\alpha}$ (globulin) inhibitor H4, inter-${\alpha}$-trypsin inhibitor heavy chain H4 precursor, human zinc-${\alpha}$-2-glycoprotein chain B, PRO2619, PRO02044, and complex-forming glycoprotein HC were new proteins that have not been previously reported in HFF using proteomic methods. Additionally, we identified alloalbumin venezia for the first time from trichloroacetic acid (TCA)-precipitated HFF. These HFF proteins could serve as new biomarkers for important human reproductive processes.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.413.2-413.2
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• 2002

Paclitaxel is an antitumor agent with poor water solubility and its pharmacokinetics are nonlinear. Cremophor EL. a surfactant used in the formulation of paclitaxel. may cause adverse effects. New solubilizer(Aceporol 460) was developed to reduce side effects of Cremophor EL and to increase the effect of drug as surfactant used in the intravenous micelle formulation of anticancer drug paclitaxel. We studied easy, rapid quantitative determination of Aceporol 460 in mouse plasma samples. which was achieved by complexation of the compound with the Coomassie brilliant blue G-250 dye in protein-free extracts. (omitted)

• Korean Journal of Ecology and Environment
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• v.35 no.3 s.99
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• pp.232-236
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• 2002

In an effort to uncover the reproduction of Korean brook perch Coreoperca herzi testis anatomy and sperm morphology were studied. Fish samples were collected in the Sooypcheon river from May to October 2001. White-colored testes have wedgeshaped external morphology, and developed symmetrically in the dorsal cavity of the trunk. Isogenetic germ cells developed in the cyst located in seminiferous lobule. Each lobule showed significant asynchrony in the spermatogenic stage of the cyst. Sperm was 43 ${\mu}$m in length. The round head was 2.2 ${\mu}$m long. The middle piece developed beneath the head was 0.5 ${\mu}$m long. Tail was 40 ${\mu}$m in length. Coomassie brilliant blue (CBB) gave rise the intense staining in the apex of sperm head and middle piece, suggesting the possible development of acrosome.

• Journal of Sericultural and Entomological Science
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• v.31 no.2
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• pp.82-90
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• 1989

Antheraea yamamai vitellin was purified from matured eggs by polyacrylamide gel electrophoresis for characterization of its biochemical properties : molecular weight, sugar and lipid composition, amino acid composition and electron microscopic morphology, etc. 1. A yamamai vitellin was composed of two subunits, large and small, showing different mobility in SDS-polyacrylamide gel electrophoresis. 2. The molecular weight of the vitellin was estimated to be approximately 450,000 dalton and the large and small subunits were 174,000 dalton and 44,000 dalton, respectively. 3. The vitellin seemed to be a glycolipoprotein since it showed a positive reaction to coomassie brilliant blue, sudan black B and PAS staining. Both subunits were similiar in this aspect. 4. Lipid of the witellin reveraled several different types including saturated lipids. 5. When the vitellin was incubated at 7$0^{\circ}C$ for 60 minites its apoprotein still cross-reacted to the specific antiserum to the native vitellin. Its sugar components were also detected by PAS staining, but its lipid portion was not detected by sudan black B staining. 6. Its amino acid composition was similar to that of other insects, but its glycine content was peculiarly very high. 7. The vitellin molecule was spherical in shape with a diameter of 14$\pm$0.8nm by negatively.

• Journal of the Korean Wood Science and Technology
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• v.33 no.4 s.132
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• pp.45-52
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• 2005

The manganese peroxidase (mnp5) from white-rot fungus Phanerochaete chrysosporium has been heterologously expressed in the methylotrophic yeast Pichia pastoris. The majority of the rMnP5 (recombinant MnP5) produced by P. pastoris exhibited an approximate molecular mass 45 kDa considerably larger than that of the predicting mnp5 due to two glycosylation sites of mnp5. After site direct mutation treatment, the effect of N-linked hyperglycosylation was examined by enzyme activity. Analysis by sodium dodesyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Coomassie Brilliant Blue (CBB) staining revealed a major protein band with a molecular mass of 37 kDa. Enzyme activity of M-rMnP5 (mutant recombinant MnP5) was similar to that of rMnP5, indicating that hyperglycosylation did not affect the active site. In this work, active mnp5 was successfully expressed in P. pastoris, suggesting that P. pastoris has potential capability of producing active heme-containing proteins.