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Determining Zebrafish Epitope Reactivity to Commercially Available Antibodies
- Source :
- Zebrafish. 14(4)
- Publication Year :
- 2017
-
Abstract
- Antibodies raised against mammalian proteins may exhibit cross-reactivity with zebrafish proteins, making these antibodies useful for fish studies. However, zebrafish may express multiple paralogues of similar sequence and size, making them difficult to distinguish by traditional Western blot analysis. To identify the zebrafish proteins that are recognized by an antimammalian antibody, we developed a system to screen putative epitopes by cloning the sequences between the yeast SUMO protein and a C-terminal 6xHis tag. The recombinant fusion protein was expressed in Escherichia coli and analyzed by Western blot to conclusively identify epitopes that exhibit cross-reactivity with the antibodies of interest. This approach can be used to determine the species cross-reactivity and epitope specificity of a wide variety of peptide antigen-derived antibodies.
- Subjects :
- 0301 basic medicine
Recombinant Fusion Proteins
Blotting, Western
Sequence Homology
Biochemistry
Epitope
Antibodies
03 medical and health sciences
Western blot
Antibody Specificity
Genetics
medicine
Animals
Humans
Reactivity (chemistry)
0501 psychology and cognitive sciences
Histidine
050102 behavioral science & comparative psychology
Amino Acid Sequence
Molecular Biology
Peptide sequence
Zebrafish
Linear epitope
biology
medicine.diagnostic_test
Chemistry
Receptor, Melatonin, MT2
Receptor, Melatonin, MT1
05 social sciences
TechnoFish—Methods
Zebrafish Proteins
biology.organism_classification
Fusion protein
030104 developmental biology
Epitope mapping
biology.protein
Small Ubiquitin-Related Modifier Proteins
Animal Science and Zoology
Antibody
Epitope Mapping
Biotechnology
Developmental Biology
Subjects
Details
- ISSN :
- 15578542
- Volume :
- 14
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- Zebrafish
- Accession number :
- edsair.doi.dedup.....c16ea877b042fa9f88ca17e6338d4738