• Bulletin of the Korean Chemical Society
• /
• v.23 no.1
• /
• pp.29-34
• /
• 2002

The microphase adsorption - spectral correction (MPASC) technique was described and applied to study the interaction of fluorescein isothiocyanate (FITC) with cetyl trimethylammonium bromide (CTAB). The synergism mechanism of micelle was analyzed and discussed. The aggregation of FITC on CTAB obeys the Langmuir monolayer adsorption. Results showed that the monomer and micellar aggregate is $FITC-CTAB_3$ and $(FITC-CTAB_3)_{29}$, respectively at pH 9.62. The adsorption constant of the CTAB-FITC aggregate was determined to be $2.10{\times}10^5$. Interestingly, it was observed for the addition of an anionic surfactant to desorb FITC from its CTAB aggregate and this has been applied to the quantitative determination of trace amounts of anionic detergent (AD) in sewage.

• Journal of Photoscience
• /
• v.10 no.3
• /
• pp.237-240
• /
• 2003

The tetradecameric peptide (K47-K60) near the NH$_2$-terminal region of epithelial-cell adhesion molecule (Ep-CAM) was chosen as antigenic site and a polyclonal antibody was generated, which could recognize Ep-CAM from the mouse colon tissue or the colon cancer cell, CT-26, in Western blot analysis. Then, the fluorescein isothiocyanate (FITC), a fluorescence dye, was conjugated with the affinity purified Ep-CAM antibody using thiocyanate and the amino groups of FITC and antibody, respectively. The molar ratio of FITC to antibody was estimated approximately 1.86 to 1.00 by measuring the optical densities at 492 nm and 280 nm. Ep-CAM antibody-FITC conjugate was then used for immunohistochemistry of the CT-26 cells. Judging from the shapes formed by fluorescence, the Ep-CAM antibody could delivered FITC to the surface of cells in which Ep-CAM was expressed. This result implies that Ep-CAM antibody could be also used for the tissue-specific delivery of the photosensitizer to the target protein via antigen-antibody interaction.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.14 no.8
• /
• pp.4106-4110
• /
• 2013

Apolipophorin-III (apoLp-III) was isolated and purified from the last larval hemolymph of Galleria mellonella by the KBr gradient ultracentrifugation and gel chromatography (Sephadex G-100). In this paper, we examined that apoLp-III is taken up into the last larval fat bodies in Galleria mellonella. The last larval fat body tissues were incubated at room temperature for 30 min with fluorescein isothiocyanate (FITC)-labeled apoLp-III (FITC-apoLp-III). Fluorescein microscopy and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) revealed that the last larval fat body tissues internalize FITC-apoLp-III. The results show that the apoLp-III is taken up by the last larval fat body.

• ALGAE
• /
• v.24 no.1
• /
• pp.39-45
• /
• 2009

The conjugation processes of a filamentous freshwater green alga Spirogyra varians were examined using FITC-lectins. Conjugation comprised five steps: 1) aligning with adjacent filaments, 2) formation of conjugation protru-sion (papilla), 3) fusion of the protrusions, 4) formation of conjugation tube,and 5) formation of zygotes. Three lectins, ConA, RCA and UEA, showed considerable labeling during the progression of conjuation. FITC-ConA labeled the surfaces of filaments throughout the whole conjugation processes. FITC-RCA labeling was observed at the conjugation protrusions only after the papilla formation. Strong labeling continued until formationg of zygotes at the contacting area where the conjugation tube developed, but no labeling was detected on the surface of vegetative filaments. The labeling decreased gradually over time and disappeared when zygotes were formed. FITC-UEA showed similar labeling pattern with FITC-RCA except that weak labeling remained after zygote formation. Inhibition experiments using RCA, UEA which are complementary to sugars L-fucose and D-galactose, showed considerable decrease of conjugation (<32% vs. 70% in control). These results suggested that the lectin-carbohydrate recognition system might be involved in the conjugation of spirogyra varians.

• Korean Journal of Plant Tissue Culture
• /
• v.21 no.6
• /
• pp.363-367
• /
• 1994

In an effort to investigate the role of calmodulin (CaM) as a modulating molecule in the signal transduction system in plant cells, we established methods for introduction of purified CaM into cultured soybean cells. CaM was purified from bovine testis, and was labelled with fluorescein isothiocyanate (FITC). Suspension -cultured cells were healed with saponin (0.1 mg/mL) to permeabilize the plasma membrane and coincubated with FITC-CaM complex. Saponin pretreatment was found to increase the fluorescence in the suspension cultured cells, indicating that the FITC-CaM complex could be incorporated into the cytoplasm. Optimal conditions for introducing FITC-CaM complex into protoplasts by electroporation were established with various electric pulses. With increasing field strength, the fluorescence in the protoplase was increased, while the viability of the protoplase decreased. FITC-CaM complex was successfully introduced into the protoplasts by electroporation and the amount of FITC-CaM complex in the protoplase was estimated.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.17 no.10
• /
• pp.199-203
• /
• 2016

Apolipophorin-III (apoLp-III) was isolated and purified from the last larval hemolymph of Galleria mellonella by KBr gradient ultracentrifugation and gel chromatography (Sephadex G-100). After KBr gradient ultracentrifugation, the lipophorin-free fractions were used as the samples for gel chromatography. The purity of the finally purified apoLp-III was confirmed by SDS-PAGE after gel chromatography. In this study, we found that apoLp-III is taken up into the adult testes in Galleria mellonella. The testes were dissected from day-1 or -2 adults in cold Ringer's solution and used for tissue culture. The protein moiety of apoLp-III was labeled with FITC dissolved in dimethyl sulfoxide (DMSO) at room temperature under conditions of continuous stirring for 1 h. The FITC-labeled apoLp-III was purified with a Sephadex G-25 PD-10 column. The tissues of the adult testes were incubated at room temperature for 30 min with fluorescein isothiocyanate (FITC)-labeled apoLp-III. Fluorescein microscopy and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) revealed that the adult testis tissues internalize the FITC-labeled apoLp-III. The results showed that apoLp-III is taken up by the adult testes.

• Journal of Pharmaceutical Investigation
• /
• v.38 no.6
• /
• pp.357-363
• /
• 2008

Penetration rate of large molecule through skin is very low due to the barrier effect of stratum corneum. Novel microneedle treatment device with roll was designed for transdermal delivery of large molecular drugs such as vaccine and protein drugs. The permeation rates of FITC labelled bovine serum albumin (FITC-BSA) as a model protein were determined using modified Franz diffusion cell and hairless mouse skin which were treated by hydrogel or solution containing FITC-BSA. Fluorescent spectrophotometer was used to analyze the concentration of FITC-BSA. Microscope using fluorescent filter was used to capture the image and location of FITC-BSA in the skin. We confirmed that permeation rate of BSA was increased with the treatment by microneedle and was increased by the increasing frequency of treatment. Furthermore, the permeation rate observed from hydrogel treated skin was significantly higher than that from solution treated skin.

• Animal cells and systems
• /
• v.5 no.1
• /
• pp.51-57
• /
• 2001

We have previously shown that oocyte maturation is induced by an immobilized progesterone, progesterone-3-carboxymethyloxime - bovine serum albumin conjugate (P-BSA) in Rana dybowskii. In this study, we confirmed the maturation inducing activity of P-BSA on Xenopus oocyte and examined the binding character of the immobilized progesterone on the surface of Xenopus oocytes after removal of the vitelline layer. P-BSA induced maturation of Xenopus oocytes but E-BSA failed to do so as observed in Rana. Binding of the immobilized progesterone, fluorescein isothiocyanate-labeled progesterone-3-0-carboxymethyloxime-BSA (P-BSA-FITC) on the devitellined oocytes surface was examined by fluorescence confocal microscopy. The binding affinity of P-BSA-FITC to the devitellined oocyte was higher than that of estrogen-BSA-FITC (E-BSA-FITC) or testosterone-BSA-FITC (T-BSA-FITC). The binding disappeared in the presence of excess free progesterone but not in the presence of free estrogen. Maximum binding occurred after two-hours of incubation with P-BSA-FITC at pH 7.5. Stronger binding occurred in oocytes at stage Vl than stage IV, and in vitro treatment of hCG enhanced the binding. Taken together, these results suggest that a specific receptor for progesterone exists on the plasma membrane of Xenopus oocytes and that progesterone acts initially on this putative receptors and triggers generation of membrane-mediated second messengers during the early stage of oocyte maturation In amphibians.

• ALGAE
• /
• v.25 no.4
• /
• pp.197-204
• /
• 2010

Cytoskeletal changes were observed during cell division of the green alga Zygnema cruciatum using flourescein isothiocynate (FITC)-conjugated phallacidin for F-actin staining and FITC-anti-$\alpha$-tubulin for microtubule staining. Z. cruciatum was uninucleate with two star-shaped chloroplasts. Nuclear division and cell plate formation occurred prior to chloroplast division. Actin filaments appeared on the chromosome and nuclear surface during prophase, and the F-actin ring appeared as the cleavage furrow developed. FITC-phallacidin revealed that actin filaments were attached to the chromosomes during metaphase. The F-actin ring disappeared at late metaphase. At telophase, FITC-phallacidin staining of actin filaments disappeared. FITC-anti-$\alpha$-tubulin staining revealed that microtubules were arranged beneath the protoplasm during interphase and then localized on the nuclear region at prophase, and that the mitotic spindle was formed during metaphase. The microtubules appeared between dividing chloroplasts. The results indicate that a coordination of actin filaments and microtubules might be necessary for nuclear division and chromosome movement in Z. cruciatum.

• Animal cells and systems
• /
• v.9 no.2
• /
• pp.65-73
• /
• 2005

In this study, we found that sperm ball of Urechis unicinctus consisted of a somatic cell and spermatogenic cells. After separation from the sperm ball, individual spermatid floated freely in the coelomic fluid and differentiated into a mature sperm. Because of many nuclear vacuoles, spermatid nucleus was observed to be heterogeneous. Later, the spermatid nucleus condensed into the homogeneous round nucleus of the mature sperm. Perinuclear microtubules could be seen but did not seem to be organized into manchette microtubules. To understand the nature of nuclear condensation during spermiogenesis, the sperm and spermatids (spermiogenic cells) were treated with FITC-phalloidin, or anti-actin-FITC, or labeled with antiactin immunogold particles (AAIP; 10 nm) followed by transmission electron microscopy or confocal laser scanning microscopy. The anti-actin-FITC and FITC-phalloidin reactions occurred distinctly in the nuclei of both spermiogenic cells. FITC-phalloidin reacted more intensely with acrosomes. The AAIP were incorporated mainly into nuclei of both cells sometimes showing local distribution in the nucleus. Nuclear vacuoles of spermatids disappeared progressively with condensation of the nucleus, as the number of incorporated $AAIP/{\mu}m^2$ increased. These results suggest that nuclear actin microfilaments might be closely related to nuclear condensation.