Abstract: Shadoo protein (Sho) was described in 2003 as the newest member of Prion protein superfamily [1]. Sho has similar structural motifs like prion protein (PrP) that is known for its central role in transmissible spongiform enchephalopathies. Although a great number of functions have been proposed, the exact physiological function of PrP is not known yet. Investigation of the function and localization of Sho may help us to understand the function of the Prion protein superfamily. Analyzing the subcellular localization of YFP-tagged forms of Sho, we detected the protein in the plasma membrane and in the nucleus of various cell lines. To reveal the localization of the endogenous protein we generated antibodies against Shadoo as well as employed commercially available anti-Shadoo antibodies: i) EG62 anti-mouse Shadoo antibody generated by Eurogentec Ltd.; ii) S-12 anti-human Shadoo antibody by Santa Cruz Biotechnology Inc.; iii) R-12 anti-mouse Shadoo antibody by Santa Cruz Biotechnology Inc.; iv) SPRN antibody against human Shadoo by Abgent Inc. We carried out immunocytochemistry on non-transfected HeLa, Zpl 2-1, Zw 3-5, GT1-1, GT1-7 and SHSY5Y cells as well as on YFP-Sho, Sho-YFP, and YFP-GPI transfected HeLa cells. Their specificity (in antibody-peptide competition assay) and co-localization (with the YFP signal) were assessed.
Abstract: Ethanol is generally used as a therapeutic reagent against Hepatocellular carcinoma (HCC or hepatoma) worldwide, as it can induce Hepatocellular carcinoma cell apoptosis at low concentration through a multifactorial process regulated by several unknown proteins. This paper provides a simple and available proteomic strategy for exploring differentially expressed proteins in the apoptotic pathway. The appropriate concentrations of ethanol required to induce HepG2 cell apoptosis were first assessed by MTT assay, Gisma and fluorescence staining. Next, the central proteins involved in the apoptosis pathway processs were determined using 2D-PAGE, SDS-PAGE, and bio-software analysis. Finally the downregulation of two proteins, AFP and survivin, were determined by immunocytochemistry and reverse transcriptase PCR (RT-PCR) technology. The simple, useful method demonstrated here provides a new approach to proteomic analysis in key bio-regulating process including proliferation, differentiation, apoptosis, immunity and metastasis.
Abstract: The purpose of research was to know the role of
immunogenic protein of 49 kDa from V.alginolyticus which capable
to initiate molecule expression of MHC Class II in receptor of
Cromileptes altivelis. The method used was in vivo experimental
research through testing of immunogenic protein 49 kDa from
V.alginolyticus at Cromileptes altivelis (size of 250 - 300 grams)
using 3 times booster by injecting an immunogenic protein in a
intramuscular manner. Response of expressed MHC molecule was
shown using immunocytochemistry method and SEM. Results
indicated that adhesin V.alginolyticus 49 kDa which have
immunogenic character could trigger expression of MHC class II on
receptor of grouper and has been proven by staining using
immunocytochemistry and SEM with labeling using antibody anti
MHC (anti mouse). This visible expression based on binding between
epitopes antigen and antibody anti MHC in the receptor. Using
immunocytochemistry, intracellular response of MHC to in vivo
induction of immunogenic adhesin from V.alginolyticus was shown.