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Studying Protein Function and the Role of Altered Protein Expression by Antibody Interference and Three-dimensional Reconstructions
Published on: April 21, 2016
Development and characterization of cholinephosphotransferase antibody
D Chatterjee1, S Mukherjee, S K Das
1Department of Biochemistry, Meharry Medical College, 1005 D.B. Todd Boulevard, Nashville, Tennessee 37210, USA.
Biochemical and Biophysical Research Communications
|July 27, 2001
Summary
Researchers developed specific antibodies against carnitine palmitoyltransferase (CPT) using synthetic peptides. These antibodies successfully recognized and inhibited CPT activity in guinea pig liver fractions, confirming their specificity.
Area of Science:
- Biochemistry
- Molecular Biology
- Immunology
Background:
- Carnitine palmitoyltransferase (CPT) is crucial for fatty acid metabolism.
- Specific antibodies are needed to study CPT's function and regulation.
Purpose of the Study:
- To generate and characterize antibodies against CPT.
- To confirm the specificity and inhibitory function of these antibodies.
Main Methods:
- Antibodies were generated in rabbits against synthetic peptides from yeast and guinea pig CPT cDNA.
- Antibody titers were measured using dot blot and ELISA.
- Antibody specificity was confirmed by Western blot analysis of guinea pig liver mitochondrial and microsomal proteins.
- CPT activity inhibition was assessed in a dose-dependent manner.
Main Results:
- High antibody titers were achieved against CPT peptides.
- Western blot analysis showed the antibody recognized a single protein band of approximately 46 kDa in guinea pig liver fractions.
- The generated antibody demonstrated dose-dependent inhibition of CPT activity in both mitochondrial and microsomal fractions.
Conclusions:
- The study successfully generated specific antibodies against carnitine palmitoyltransferase (CPT).
- These antibodies are suitable tools for studying CPT in biological samples.
- The antibodies can be used to investigate CPT's role in cellular processes and potentially in disease states.
