Production and characterization of polyclonal antibody against a synthetic peptide from β-actin protein

Authors

  • Mohadeseh Naghi Vishteh Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
  • Nazila Amini Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran 2 Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
  • Negah Ahmadvand Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
  • Omid Zarei Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran
  • Reza Hadavi Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
Abstract:

Objective(s):Antibodies against actin, as one of the most widely studied structural and multifunctional housekeeping proteins in eukaryotic cells, are used as internal loading controls in western blot analyses. The aim of this study was to produce polyclonal antibody against a synthetic peptide derived from N-terminal region of β-actin protein to be used as a protein loading control in western blot and other assay systems. Materials and Methods: A synthetic peptide derived from β-actin protein was designed and conjugated to Keyhole limpet hemocyanin (KLH (and used to immunize a white New Zealand rabbit. The antibody was purified from serum by affinity chromatography column. The purity of the antibody was determined by SDS-PAGE and its ability to recognize the immunizing peptide was measured by ELISA. The reactivity of the antibody with β-actin protein in a panel of different cell lysates was then evaluated by western blot. In addition, the reactivity of the antibody with the corresponding protein was also evaluated by Immunocytochemistry and Immunohistochemistry in different samples. Results: The antibody could recognize the immunizing peptide in ELISA. It could also recognize            β-actin protein in western blot as well as in immunocytochemistry and immunohistochemistry. Conclusion: Our data suggest that this antibody may be used as an internal control in western blot analyses as well as in other immunological applications such as ELISA,immunocytochemistry and immunohistochemistry.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

production and characterization of polyclonal antibody against a synthetic peptide from β-actin protein

objective(s):antibodies against actin, as one of the most widely studied structural and multifunctional housekeeping proteins in eukaryotic cells, are used as internal loading controls in western blot analyses. the aim of this study was to produce polyclonal antibody against a synthetic peptide derived from n-terminal region of β-actin protein to be used as a protein loading control in western ...

full text

peptide-based polyclonal antibody production against p110 protein of mycoplasma genitalium

mycoplasma genitalium (m.genitalium) is a sexually transmitted pathogen. detection of this microorganism in clinical specimens by culture is rather difficult and time consuming. the aim of this study was to produce polyclonal antibody against a synthetic peptide from p110 protein of m.genitalium in order to develop a diagnostic tool for detection of this microorganism in clinical specimens. a s...

full text

Preparation and Characterization of a Polyclonal Antibody against Human Actin Filament-Associated Protein-120 kD

Actin filament-associated protein-120kD (AFAP-120) is an alternatively spliced isoform of actin filament-associated protein-110kD (AFAP-110) and contains an additional neuronal insert (NINS) fragment in addition to identical domains to the AFAP-110. Unlike AFAP-110 widely expressed in tissues, AFAP-120 is specifically expressed in the nervous system and plays a role in organizing dynamic actin ...

full text

Preparation and Characterization of a Polyclonal Antibody against Brominated Protein

(Di)bromotyrosine is formed by the specific reaction of eosinophil peroxidase and can be used as an eosinophil activation marker. In the present study, an antibody for (di)bromotyrosine in proteins was prepared to investigate the pathogenesis of eosinophil-related diseases such as allergic responses. A rabbit polyclonal antibody was raised against brominated keyhole limpet hemocyanin. The speci...

full text

Peptide based polyclonal antibody production against bovine rotavirus non structure protein4 (NSP4)

The rotavirus nonstructural protein 4 (NSP4), is a multi functional protein that play key role in both viral morphogenesis and cytopathic effect associated with cell death. However, the complete biological effect of NSP4 remains to be clarified. Since to obtain further knowledge about this protein there is a need for recognizing antibody and there is no commercial antibody against this protein,...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 17  issue 6

pages  396- 400

publication date 2014-06-01

By following a journal you will be notified via email when a new issue of this journal is published.

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023