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STIMULATOR OF NEUROTROPIC EFFECTS: DETERMINING THE MECHANISM OF ACTION OF THE MS-818 COMPOUND THROUGH PROTEIN IDENTIFICATION BY AFFINITY CHROMATOGRAPHY AND SDS-PAGE

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Date Issued:
2011
Abstract/Description:
The MS-818 compound is used in the proliferation process of neuronal cells and many biological activities that accompany this process such as astrocyte differentiation, inhibition of neuronal apoptosis, and fraction repairs. We do know the effects of this compound, but the mechanism of action remained uncertain until now. To determine the pathway of this compound, NT2 cells were cultured and lysed to isolate the proteins. Affinity Chromatography was performed in order to immobilize the MS-818 compound to a Hi-Trap NHS column. The NT2 protein sample was injected through the column and eluted with a MS-818 concentrated, high salt content elution buffer. SDS-PAGE was then performed to isolate the proteins that bound to MS-818. The gel was visualized using Coomassie Blue. The results indicate that there are two proteins associated in the mechanism of this compound. A standard protein marker ranging from 10 kDa to 250 kDa was used to compare the bands. The findings indicate that one of the protein bands is slightly less than 250kDa and the other is between 50-75 kDa. When the proteins are confirmed by mass spectrometry sequencing, this will help to promote this compound as a drug candidate.
Title: STIMULATOR OF NEUROTROPIC EFFECTS: DETERMINING THE MECHANISM OF ACTION OF THE MS-818 COMPOUND THROUGH PROTEIN IDENTIFICATION BY AFFINITY CHROMATOGRAPHY AND SDS-PAGE.
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Name(s): Dass, Charlene, Author
Sugaya, Kiminobu, Committee Chair
University of Central Florida, Degree Grantor
Type of Resource: text
Date Issued: 2011
Publisher: University of Central Florida
Language(s): English
Abstract/Description: The MS-818 compound is used in the proliferation process of neuronal cells and many biological activities that accompany this process such as astrocyte differentiation, inhibition of neuronal apoptosis, and fraction repairs. We do know the effects of this compound, but the mechanism of action remained uncertain until now. To determine the pathway of this compound, NT2 cells were cultured and lysed to isolate the proteins. Affinity Chromatography was performed in order to immobilize the MS-818 compound to a Hi-Trap NHS column. The NT2 protein sample was injected through the column and eluted with a MS-818 concentrated, high salt content elution buffer. SDS-PAGE was then performed to isolate the proteins that bound to MS-818. The gel was visualized using Coomassie Blue. The results indicate that there are two proteins associated in the mechanism of this compound. A standard protein marker ranging from 10 kDa to 250 kDa was used to compare the bands. The findings indicate that one of the protein bands is slightly less than 250kDa and the other is between 50-75 kDa. When the proteins are confirmed by mass spectrometry sequencing, this will help to promote this compound as a drug candidate.
Identifier: CFH0004076 (IID), ucf:44807 (fedora)
Note(s): 2011-08-01
B.S.
Medicine, Dept. of Molecular Biology and Microbiology
Undergraduate
This record was generated from author submitted information.
Subject(s): MS-818
SDS-PAGE
Affinity Chromatography
Persistent Link to This Record: http://purl.flvc.org/ucf/fd/CFH0004076
Restrictions on Access: public
Host Institution: UCF

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