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Gyurng-Soo Yoo,Chung Ki Sung 한국생약학회 1986 생약학회지 Vol.17 No.2
For the immunoassay of ginseng sapogenins, the ligands with which panaxadiol and panaxatriol could be determined together and separately were synthesized. For the total assay of panaxadiol and panaxatriol, panaxatriol-6-hemisuccinate was synthesized. For the separate assay, panaxadiol-3-hemisuccinate and panaxatriol-3-hemisuccinate were also synthesized.
Direct Blue 71 staining of proteins bound to blotting membranes
Hong, Hee Youn,Yoo, Gyurng Soo,Choi, Jung Kap 전남대학교 약품개발연구소 2000 약품개발연구지 Vol.9 No.1
A sensitive staining method for protein blots using Direct Blue 71 is described. It is based on the selective binding of dye molecules proteins in acidic solution and produces blush violet colored bands. It is a simple and rapid procedure, involving only staining and rinsing steps that occur within 7 min. The sensitivity of this method is 5-10 ng of protein on nitrocellulose (NC) and 10-20 ng, on polyvinylidene difluoride (PVDF), which is tenfold better Than that of the, commonly used Ponceau S staining. Moreover, the staining is reversible for subsequent immunostaining, without impairing immunoreactivity. To remove the dye from the developed bands, changes in pH and hydrophobicity of the solvent are required. Due to its sensitivity, rapidity, simplicity, and tow cost, this stain may be more practical than other dye-used stains or metal-based stains for routine laboratory purposes.
Hong, Hee Youn,Yoo, Gyurng Soo,Choi, Jung Kap 전남대학교 약품개발연구소 1998 약품개발연구지 Vol.7 No.1
In the present study, we examined effects of ginseng saponins (giasenosides) on pp60^(c-src) protein tyrosine kinase (PTK) activity, intracellular calcium concentration ([Ca^(2+)]_i)₁ and cell proliferation in NIH3T3 cells. Eight different giasenosides [ginsenoside-Rb, (G-Rb,) -Rb₂, -Rc₁ -Rd, -Re, -Rf, -Rg₁, -Rg₂] and ginseng total saponin (GTS) were used for these experiments. All ginsenosides and GTS tested stimulated the activation of pp60^(c-src) kinase, and especially G-Rb₁, -Rd, -Rg₁, and -Rg₂ showed a higher stimulatory effect than others at 16.7 ㎍/㎖ of ginsenosides with a 18 hr-incubation, increasing the activity by 4.5. 3.5, 3.5, and 3.0-fold, respectively, over that of untreated control. In addition, both G-Rd and -Rg₂ increased [Ca^(2+)]_i to 202 and 334 nM, respectively, about 2-3-fold above the basal level within 7min at 250 ㎍/㎖ of ginsenosides. The increases of [Ca^(2+)]_i]; were eliminated by pretreatment of EGTA, an extracellular calcium chelator, suggesting that they result from an influx of calcium ion from extracellular medium rather than an efflux from intracellular calcium store, endoplasmic reticulum (ER). All ginsenosides studied enhanced cell proliferation to 1.2-1.4-fold over that of untreated control at 5∼250 ㎍/㎖ of concentrations. Interestingly the promotion of cell proliferation by ginsenosides corresponded with the activation of c-src kinase which is as early step is the mitogenic signaling cascade. Taken together, we suggest that some ginsenosides may lead to cell proliferation via the activation of cellular signal transduction pathway involving pp60^(c-src) kinase.
Hong, Hee-Youn,Yoo, Gyurng-Soo,Choi, Jung-Kap The Korean Society of Ginseng 1998 Journal of Ginseng Research Vol.22 No.2
In the present study, we examined effects of ginseng saponins (ginsenosides) on pp60c-src protein tyrosine kinase (PTK) activity, intracellular calcium concentration ([$Ca^{2+}$]i), and cell proliferation in NIH3T3 cells. Eight different ginsenosides [ginsenoside-Rb1 (G-$Rb_1$), -$Rb_2$, -Rc, -Rd, -Re, -Rf, -$Rg_1$, -$Rg_2$) and ginseng total saponin (GTS) were used for these experiments. All ginsenosides and GTS tested stimulated the activation of $pp60^{c-src}$ kinase, and especially G-$Rb_1$,-Rd,-$Rg_1$, and -$Rg_1$ showed a higher stimulatory effect than others at 16.7 $\mu\textrm{g}$/ml of ginsenosides with a 18 hr-incubation, increasing the activity by 4.5, 3.5, 3.5, and 3.0-fold, respectively, over that of untreated control. In addition, both G-Rd and -$Rg_2$)Rg2 increased ($Ca^{2+}$), to 202 and 334 nM, respectively, about 2-3-fold above the basal level within 7min at 250 $\mu\textrm{g}$/yml of ginsenosides. The increases of ($Ca^{2+}$), were eliminated by Pretreatment of EGTA, an extracellular calcium chelator, suggtasting that they result from an influx of calcium ion from extracellular medium rather than an efflux from intracellular calcium store, endoplasmic reticulum (ER). All ginsenosides studied enhanced cell proliferation to 1.2-1.4-fold over that of untreated control at 5~250 $\mu\textrm{g}$/ml of concentrations. Interestingly the promotion of cell proliferation by ginsenosides corresponded with the activation of c-src kinase, which is an early step in the mitogenic signaling cascade. Taken together, we suggest that some ginsenosides may lead to cellProliferation via the activation of cellular signal transduction Pathway involving $pp60^{c-src}$ kinase.
Improved Phosphotyrosine Analysis by TLC and HPLC
Song, Young-Me,Yoo, Gyurng-Soo,Lee, Seung-Ki,Choi, Jung-Kap The Pharmaceutical Society of Korea 1993 Archives of Pharmacal Research Vol.16 No.2
We describe here the conditions of thin layer chromatography (TLC) and high pressures liquid chromatography (HPLC) to improve the analytical method of phosphotyrosine (p-Tyr) in biological sample. TLC was performed on silica plate with the mixture of propanol and water (2.1 : 1 v/v) as a mobile phase and $R_1$ values were 0.42, 0.39 and 0.33 for phosphotyrosine, phosphothreonine and phosphoserine, respectively. HPLC was performed on $NH_2$ column with a mobile phase of potassium biphosphate solution by UV deterction at 192 nm. The optimum condition of HPLC was obtained at 0.01 M, pH 4.5 with a clear separation within 12 min. These procedures have been applied to the analysis of phosphotyrosine obtained from tyrosine-phosphorylated enolase. Both TLC and HPLC methods were suitable to analyze tyrosine-phosphorylated protein without being affected by contaminants from hydrolysates.