PMID: 9438067Jan 23, 1998Paper

Preparation and purification of antisera against different regions or isoforms of beta-amyloid precursor protein

Brain Research. Brain Research Protocols
A JenA J Patel

Abstract

We describe a procedure for the production and peptide affinity purification of polyclonal antisera against synthetic peptides representing different domains of beta-amyloid precursor protein (APP). Rabbits were immunised with keyhole limpet haemocyanin coupled to synthetic peptides representing the amino-terminal APP18-32, Kunitz-type protease inhibitor (KPI) region APP301-316, the A beta region APP670-686, and the carboxy-terminal APP756-770 of APP770 for the production of antisera anti-AP-1, anti-AP-2, anti-AP-4 and anti-AP-5, respectively. Each antiserum was purified to specific antibody using the respective cognate peptides immobilised on affinity columns as ligand, using the 1-ethyl-3-(dimethylaminopropyl)carbodiimide-diaminodipropylamine method. Purified antibodies of these four antisera were highly specific and in enzyme-linked immunosorbent assays (ELISA) reacted only to the corresponding peptide. These purified antisera have been used in Western blot, immunohistochemical and immunoprecipitation techniques to facilitate the understanding of the regulation of APP and amyloid beta-protein (A beta). The A beta is formed by proteolysis of APP, and its deposition leading to the formation of senile plaques in the brain is co...Continue Reading

References

Feb 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·A TamaokaD J Selkoe
Jan 1, 1992·Journal of Neuropathology and Experimental Neurology·B T HymanD Schenk
Nov 1, 1991·Brain Research·T KawarabayashiS Hirai
Jul 8, 1991·Neuroscience Letters·J P AndersonN K Robakis
Feb 1, 1990·Analytical Biochemistry·M A GillesC L Borders
Feb 15, 1990·Analytical Biochemistry·Z Grabarek, J Gergely
Jan 1, 1988·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·T T Herskovits
Oct 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·D J SelkoeT Oltersdorf
Oct 1, 1987·Analytical Biochemistry·M DianoM Hirn
Dec 1, 1980·Analytical Biochemistry·P K SmithG T Hermanson
Aug 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·H Kimura, D Schubert
May 9, 1995·Proceedings of the National Academy of Sciences of the United States of America·L S HigginsB Cordell
Mar 13, 1995·Brain Research·W E MüllerK Scheuer
Jan 1, 1994·Annual Review of Neuroscience·D J Selkoe
Sep 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·D T LooC W Cotman
Apr 1, 1993·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·J R MoffettJ H Neale
Dec 1, 1996·Neurodegeneration : a Journal for Neurodegenerative Disorders, Neuroprotection, and Neuroregeneration·A J PatelH A de Silva
Jul 1, 1997·Brain Research. Molecular Brain Research·H A Rohan de SilvaA J Patel

❮ Previous
Next ❯

Citations


❮ Previous
Next ❯

Related Concepts

Related Feeds

Antibody Specificity

Antibodies produced by B cells are highly specific for antigen as a result of random gene recombination and somatic hypermutation and affinity maturation. As the main effector of the humoral immune system, antibodies can neutralize foreign cells. Find the latest research on antibody specificity here.

Alzheimer's Disease: APP

Amyloid precursor protein (APP) proteolysis is critical for the development of Alzheimer's disease, a neurodegenerative disease associated with accumulation of amyloid plaques in the brain. Here is the latest research on APP and Alzheimer's disease.

Alzheimer's Disease: Amyloid Beta

Alzheimer's disease is a neurodegenerative disease associated with the accumulation of amyloid plaques in the brain; these plaques are comprised of amyloid beta deposits. Here is the latest research in this field.