• Bulletin of the Korean Chemical Society
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• 제23권1호
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• pp.29-34
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• 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
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• 제10권3호
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• pp.237-240
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• 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.

• 한국산학기술학회논문지
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• 제14권8호
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• pp.4106-4110
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• 2013

아포리포포린-III (apoLp-III)를 꿀벌부채명나방 종령 유충 혈림프에서 KBr 농도구배 초원심분리와 겔크로마토그래피 (Sephadex G-100)를 이용하여 분리, 정제하였다. 본 연구에서는 아포리포포린-III가 꿀벌부채명나방의 종령 유충 지방체에 의해 흡수되는 지를 조사하였다. 종령 유충 지방체 조직을 FITC로 표지한 아포리포포린-III(FITC-apoLp-III)와 상온에서 30분간 배양하였다. 배양 후 형광현미경과 전기영동을 이용하여 FITC-apoLp-III가 지방체 조직으로 들어가는 지의 여부를 확인하였다. 그 결과 FITC-apoLp-III가 유충 지방체로 흡수된다는 사실을 알 수 있었다.

• ALGAE
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• 제24권1호
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• pp.39-45
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• 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.

• 식물조직배양학회지
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• 제21권6호
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• pp.363-367
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• 1994

식물 세포의 신호전달 기작에 있어 calmodulin (CaM)의 역할을 조사하기 위하여 외부기원의 CaM을 식물 세포 내로 주입하였다. 이를 위하여 bovine testis로부터 CaM을 분리, 정제하여 SDS-PAGE상에서 순수함을 확인하였고, 이를 세포 내로 주입한 후 실제 주입 여부를 확인하기 위해 정제된 CaM을 미리 fluorescein isothiocyanate (FITC)로 표식하여 사용하였다. 또한, FITC-CaM complex의 농도에 따른 fluorescent intensity를 측정하여 주입된 CaM의 농도를 결정하였다. 대두 현탁 배양 세포를 saponin (0.1 mg/mL)이 포함된 배지에서 15분간 배양하여 permeabilization시키고,FITC-CaM (1mg/mL)을 30분간 처리한 후 형광현미경으로 조사했을 때 세포질에서 강한 형광이 나타났으며, 이 결과로 외부의 calmodulin이 세포 내로 주입되었음을 확인할 수 있었다. 대두 현탁 배양 세포에 3가지의 세포벽 분해 효소를 처리하여 원형질체를 분리하였고, 이 원형질체를 FITC-CaM이 들어있는 완충용액에 넣고 electroporation 하였을때 capacitance와 field strength가 증가할수록 원형질체의 생존률은 감소하였으나, 세포 내의 fluorescent intensity는 증가하였다. 이것은 외부의 calmodulin이 electroporation에 의해 대두의 원형질체 내로 도입되었음을 의미하는 것이다.

• 한국산학기술학회논문지
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• 제17권10호
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• pp.199-203
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• 2016

아포리포포린-III (apoLp-III)를 꿀벌부채명나방 종령 유충 혈림프로부터 KBr 농도구배 초원심분리와 겔크로마토그래피(Sephadex G-100)를 이용하여 분리, 정제하였다. KBr 농도구배 초원심분리가 끝난 후, 리포포린을 제외한 분획 (lipophorin-free fractions)을 겔크로마토그래피 시료로 사용하였으며, 겔크로마토그래피를 행한후 sodium dodecyl sulfate (SDS)-전기영동으로 apoLp-III의 정제를 확인하였다. 또한, 본 연구에서는 유충 혈림프로부터 정제된 apoLp-III가 꿀벌부채명나방의 성충 정소에 의해 흡수되는 지를 조사하였다. 우화한 지 1일 또는 2일된 성충으로부터 성충 정소를 차가운 링거액에서 분리한 후 조직 배양의 시료로 사용하였다. 순수 정제한 apoLp-III를 dimethyl sulfoxide (DMSO)에 녹인 형광물질 fluorescein isothiocyanate (FITC)와 상온에서 계속 저어가면서 1시간 동안 배양하였다. FITC로 표지된 apoLp-III (FITC-labeled apoLp-III)를 Sephadex G-25 PD-10 column을 이용하여 정제하고 확인하였다. 정제된 FITC-labeled apoLp-III와 성충 정소 조직을 상온에서 30분간 배양하였다. 배양 후 형광현미경 (fluorescence Axioskop microscope)과 SDS-전기영동을 이용하여 FITC-labeled apoLp-III가 정소 조직으로 들어가는 지의 여부를 확인하였다. 그 결과 FITC-labeled apoLp-III가 성충 정소로 흡수된다는 사실을 알 수 있었다.

• Journal of Pharmaceutical Investigation
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• 제38권6호
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• pp.357-363
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• 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
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• 제5권1호
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• pp.51-57
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• 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
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• 제25권4호
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• pp.197-204
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• 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
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• 제9권2호
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• pp.65-73
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• 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.