Renewable Affinity Reagents
Type: Chemical / Small Molecules,
Keywords: High-quality antibody, Recombinant immunolabel, Recombinant monoclonal antibody (R-mAb), NeuroMabs, Affinity reagent, Nanobody (nAb), Intrabody
Renewable recombinant antibodies and affinity reagents for neuroscience research
We develop, validate, and enhance community access to a diverse toolbox of renewable recombinant antibodies and affinity reagents for neuroscience research. Extensively characterized in mammalian brain samples (rat, mouse, human).These reagents include conventional and recombinant mouse monoclonal antibodies and miniaturized ScFV derivatives, and nanobodies (single chain miniaturized antibodies). The small size, solubility, and stability of ScFvs and nAbs facilitates their functional expression in mammalian cells, allowing for their use as intrabodies to target cargo including optogenetic reports and actuators to distinct subcellular sites in neurons. Their small size also enhances the resolution of light and electron microscope imaging when they are used as immunolabels, and their penetration into intact cleared samples.
* Renewable affinity reagents to study proteins.
* Used to label, capture, and/or influence activity of specific targets in brain neurons.
* UC Davis/NIH NeuroMab Facility develops the renewable affinity reagents.
* UC Davis/NIH NeuroMab Facility provide a unique neuroscience-based approach to generating mouse monoclonal antibodies (mAbs) optimized for use in mammalian brain (NeuroMabs).
* Antibodies made available widely at low cost.
* Hybridoma cells and plasmids made widely available for researchers to produce their own antibody.
Extensively characterized in mammalian brain samples.
* Extensively charcterized for use in mammalian brain samples (rat, mouse, human).
* Used for immunohistochemical- based imaging studies of protein localization in adult, developing and pathological brain samples.
* Used for biochemical analyses of subunit composition and post-translational modifications of native brain proteins.
* Used for proteomic analyses of native brain protein networks.
* ScFVs and nanobodies can be used as genetically-encoded intrabodies to deliver cargo to specific subcellular locations in neurons.
* Mouse, Rat, Zebrafish, Xenopus, Chicken, Barn Owl.
* Renewable antibodies enhance research reproducibility.
* Recombinant antibodies are unambiguosly identified by sequence.
* Nanoscale immunolabels for better tissue penetration and resolution.
* Enhanced resolution of light and electron microscope imaging.
* Validated for use in mammalian brain.
* Criteria for immunoblots are monospecific reaction against a band similar in molecular characteristics, and absence of the band in KO samples
* Criteria for immunohistochemistry is a match to expected regional, cellular and subcellular pattern of expression, and absence of signal in KO samples
* Various distributors distribute the antibodies (finished product) generated by UC Davis/NIH NeuroMab Facility. Addgene distributes the plasmids encoding recombinant antibodies, ScFVs and nanobodies. The UC Davis MMRRC distributes the monoclonal antibody producing hybridoma cells.
* The UC Davis/NIH NeuroMab Facility was funded from 2005-2015 with a U24 cooperative grant from NINDS and NIMH, and through May 2019 by a High Impact Neuroscience Research Resource Grant (R24).
* Neuroscience Monoclonal Antibody Sequencing Initiative (NeuroMabSeq) is a concerted effort to determine and make publicly available the sequences of monoclonal antibodies valuable to neuroscience research.
* Gong et al., (2016). Developing High-Quality Mouse Monoclonal Antibodies for Neuroscience Research-Approaches, Perspectives and Opportunities. New Biotech. 33: 551-564. PMID: 26644354. doi: 10.1016/j.nbt.2015.11.007
* Andrews et al., (2019). A Toolbox of IgG Subclass-Switched Recombinant Monoclonal Antibodies for Enhanced Multiplex Immunolabeling of Brain. eLife 8:e43322. PMID: 300667360. doi: 10.7554/eLife.43322
* Dong et al., (2019). A Toolbox of Nanobodies Developed and Validated for Diverse Neuroscience Research Applications. eLife 8:e48750. PMID: 31566565. doi: 10.7554/eLife.48750
Jim Trimmer, Professor
University of California Davis School of Medicine
GRANT SUPPORT / FUNDING SOURCE
NIH U24 NS109113 “Recombinant Immunolabels for Nanoprecise Brain Mapping Across Scales”
NIH U01 NS099714 “Genetically Encoded Localization Modules for Targeting Activity Probes to Specific Subcellular Sites in Brain Neurons”
NIH R24 NS092991 “UC Davis/NIH NeuroMab Facility”
NIH R01 NS092474 “Synaptomes of Mouse and Man”