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Screening nicastrin antibodies for specificity for immunofluorescence (934.2)
Author(s) -
Deshaies Derek,
Yu Gang,
Dries Daniel
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.934.2
Subject(s) - immunofluorescence , nicastrin , antibody , antigen , biology , medicine , immunology , alzheimer's disease , disease , presenilin , pathology
Alzheimer’s disease is a type of dementia that can cause problems with memory, learning and behavior. Nicastrin is a component of the protein complex involved in the development of Alzheimer’s disease. In our lab, tissue‐specific nicastrin conditional knockout mice were developed and displayed a striking neurological phenotype. The aim of this research project, then, is to confirm the tissue‐specific knockout by using immunofluorescence. In order to ask this question, we must first identify an immunofluorescence‐compatible antibody that is specific for the nicastrin protein. This is important due to the cross reactivity of many antibodies that are thought to be anti‐nicastrin. In an effort to test for antibody specificity, researchers use pre‐blocking techniques to pre‐absorb (block) their antibodies with antigen before doing their experiment. However, this only proves that the antibody produced is specific for the antigen against which it was raised. Using wild‐type and nicastrin knockout cells, our approach will use a series of anti‐nicastrin antibodies for immunofluorescence to test their specificity towards their intended antigen, rather than staining antigens other than the one against which the antibodies were raised. Grant Funding Source : Supported by National Institute of Neurological Disorders and Stroke grant 1R01NS079796