Analysis of the expression of IBMR3 antigen in cells and tissues from rodents

Simple item page
dc.contributor.authorAlrawi, Qutaiba K. Jassim
dc.date.accessioned2014-11-03T02:08:57Z
dc.date.available2014-11-03T02:08:57Z
dc.date.issued2009
dc.descriptionMasteren_US
dc.description.abstractMonoclonal antibodies (Mab) and their fragments have been widely used for diagnostic and therapeutic purposes. IBMR3 hybridoma cells were produced in a previous study. The cell culture supernatant of the hybridoma cells was collected and stored at -20 °C freezer. The aim of this research is to investigate the expression profile of the IBMR3 antigens in different tissue of mouse, rat and cell lines HT29, 3T3 by using immunoblotting technique and immunohistochemical staining using the peroxidase technique. In this study the expression of IBMR3 antigen was evaluated in eighteen different organs taken from Balb/c mouse and Sprague Dawley rat, established cell lines, human colorectal HT29 cancer cell line and mouse fibroblast 3T3. Various mouse and rat tissues were snap- frozen and stored in liquid nitrogen. For immunoblotting six micron frozen sections of the various tissues were prepared and collected in Eppendorf tubes. The sections were then lysed in lysis buffer, and equal concentrations of lysates were run on 12% SDS-PAGE. The separated protein bands were transferred to PVDF/ polyvinylidene difluoride membrane for immunoblotting. Immunoblotting were subsequently subjected to densitometric analysis to get the value of molecular weight, peak height and raw volume of the protein band (Burnette, 1981). IBMR3Ag was expressed in mouse tissues with the highest MW (98.22) kDa and highest number (nine) of bands detected in muscle and the lowest MW in heart (75.00) kDa and liver (78.68) kDa with lowest number (2) of bands. In rat, more bands (eight) were detected in brain and kidney. While in spleen only four protein bands were detected. The highest MW was also indicated in brain (120.83 kDa), kidney (85.48 kDa) and 73.42 kDa in spleen. In both mouse and rat, highest expression of IBMR3 antigen was found in the lung and lowest in the brain. The immunohistochemical staining method was used in peroxidase technique for the investigation of the expression profile of the IBMR3 antigens in different frozen tissues from mouse and rat. The positive IBMR3 antigen were seen under light microscope as brown color, the results between weak, moderate and intense. Expressed IBMR3 antigens help to analyze the profile for molecular weight, peak height and raw volume. The different results in immunoblotting and immunohistochemistry staining gave positive expression for IBMR3 Ag in different part of the tissue sections, this shed more light on, whether the level of IBMR3 antigens are associated with pathological parameters. The results from this study suggest that the IBMR3 antigens were differentially expressed in mouse, rat tissue and cell lines. It was recommended that further research be conducted to identify and investigate the nature of the IBMR3 antigen and its potential role in tissue development.en_US
dc.identifier.urihttp://hdl.handle.net/123456789/199
dc.language.isoenen_US
dc.subjectBiological Scienceen_US
dc.subjectAntigenen_US
dc.subjectCellsen_US
dc.subjectTissuesen_US
dc.subjectRodentsen_US
dc.titleAnalysis of the expression of IBMR3 antigen in cells and tissues from rodentsen_US
dc.typeThesisen_US
Files
License bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
license.txt
Size:
1.71 KB
Format:
Item-specific license agreed upon to submission
Description:
Download
Collections
Pusat Pengajian Sains Kajihayat - Tesis