...10 HA or 10 MYC tags built into a GFP scaffold. Some GFP nanobodies  stain poorly when tagged with a Spaghetti...fusion between a reporter and a nanobody that’s encoded on a plasmid. A nanobody is a ~150 amino acid, single...previously published nanobody sequences that targeted the following antigens: GFP, RFP,  the histone protein...weak signals of fluorescent protein (i.e. GFP, RFP) Detecting GFP reconstitution across synaptic partners...protein? No worries! As more nanobodies are developed, there will be more nanobodies to turn into RANbodies....More nanobody expression plasmids can be found here Watch this video to learn more about nanobodies Additional...Monster because they recognize this GFP scaffold itself. Fortunately the GFP1 HA Spaghetti Monster RANbody doesn...

...GBPs/GFP. The new system, CRE-DOG (Cre Dependent On GFP), is activated by GFP and derivatives GFP and ...manipulate only GFP-labeled cells. Once Tang and Cepko found a description of GFP-binding nanobodies, the project...inside cells. Nanobodies binding GFP were designed in 2009. Subsequent work showed that nanobodies could be...to bind GFP. The idea of using GFP as a scaffold suddenly seemed very realistic. Envisioning GFP as a substrate...presence of GFP. These systems, known as T-DDOG and Cre-DOG, respectively, repurpose popular GFP reporter...money needed to develop new lines. Using nanobodies to create GFP scaffolds for transcriptional activation...lines expressing GFP in specific cell types to do more than just label cells. If GFP could be co-opted...

...ability of its nanobody partner, so it’s important to insert AsLOV2 at a spot in the nanobody that preserves...monitored how AsLOV2 insertion into mCherry or GFP nanobodies affected binding of their target proteins. The...about using nanobodies to control protein activity Read about RANbodies, reporter nanobody fusions Learn...the secondary nanobody toolbox for immunodetection Resources on Addgene.org Find nanobody expression plasmids...proteins, and then elute YFP.  Option #2: Opto-Nanobodies (OptoNBs) purification You skip adding a tag ...Toettcher Lab, OptoNBs are photoswitchable engineered nanobodies whose binding of their target protein is altered...have two key components: an AsLOV2 domain and a nanobody.  AsLOV2 is a light-oxygen-voltage sensing domain...

...Marcy Patrick  Listen to the nanobody toolkit podcast segment Nanobodies are single domain antibody fragments...includes a set of plasmids each containing an anti-GFP nanobody fused with a different functional element (tyrosine... different detection methods. Since the anti-GFP nanobody is able to bind fluorescent proteins derived...functionalized nanobodies described here can be used to target any protein fused with GFP, YFP, mCerulean...Zhao C, et al. RNA. 2018 PubMed PMID:29109157. Nanobody toolkit to monitor retrograde transport at the... as regular antibodies. These properties make nanobodies easier to transform and express in bacteria, ... Spiess lab developed a set of functionalized nanobodies to study retrograde transport from the cell surface...

...160374 Nanobodies- there's a simple method for that! By Michelle Cronin The benefits of nanobodies over ...antibodies. Nanobodies are easily expressed in the cytosol of E. coli, however the nanobody proteins tend...GABA-ergic interneurons. These include mDlx-GFP, hDlx-FLEX-GFP, hDlx-Flex-dTomato, mDlx-GCaMP6f, hDlx-GiDREADD-dTomato...evolution Fluorescent tools for mammalian cells Nanobody purification CRISPRi and CRISPRa system for hPSCs... conventional antibodies can’t be overlooked. Nanobodies are smaller in size, more durable, and usually...alleviates the inclusion body problem and makes nanobody production in E.coli as simple as following a ...signal peptide of MBP, followed by the desired nanobody (NB) sequence, an AviTAG, and 6xHis tag. The MBP...

...expressing a GFP-tagged version of a protein of interest. The binding of the GFP nanobody to its target... TurboID fused to a conditionally stabilized GFP-nanobody rather than a specific protein of interest. ...TurboID-nanobody protein and allows TurboID to start labeling proteins in close proximity to the GFP-tagged...GFP-tagged protein (albeit slightly offset due to the nanobody-GFP interaction). The modularity of the system provides...

...Antibodies  Secondary Nanobody Toolbox No Llamas Required - Synthetic Nanobodies Against Membrane Proteins...single-domain antibodies (sdAb), commercially known as nanobodies (Arbabi-Ghahroudi, 2017)!      Figure... single-domain antibody, under the commercial "Nanobody" name.  We’ve talked about sdAbs a number ...highlighted back in 2015, where sdAbs were used to create GFP scaffolds for transcriptional activation.  sdAbs ... Jin B, Odongo S, Radwanska M, Magez S (2023) Nanobodies: A Review of Generation, Diagnostics and Therapeutics...ijms24065994 Xu J, Xu K, Jung S, et al (2021) Nanobodies from camelid mice and llamas neutralize SARS-...resources from addgene.org  Plasmids encoding nanobodies at Addgene Sybody Generation Toolbox from Addgene...

...Variables (scFvs). Nanobodies Figure 3: Comparison of the Hcab and nanobody. Nanobodies are the variable...synthetic nanobodies, or synbodies, can be produced from plasmids, saving time and money. Nanobodies and synbodies...blot showing GFP expression in cells that have been transfected with a plasmid encoding GFP. Actin, a common...to reach. While a full antibody is ~150 kDa, a nanobody is 12-15 kDa.   Research applications for antibodies...Collection includes plasmids that express antibodies, nanobodies, and ScFvs, as well as tools for developing and...Developmental Studies Hybridoma Bank Learn more about nanobodies Watch our Antibodies 101 animation, What is an...

...sender” cells express GFP on the surface and “receiver” cells express an anti-GFP nanobody on the cell surface...fluorophore/antibody pairs. Because the anti-GFP nanobody cross-reacts with BFP, sender cells expressing...to track cell-cell interactions By Jennifer Tsang GFP-based Touching Nexus (G-baToN) is a new tool developed...surface. When the cells interact, GFP is transferred to the receiver cell, and the receiver cell turns green...expression of cytoplasmic-tdTomato and Synaptophysin-GFP. Synapse targeting GCaMP6s from the Rylan Larsen ...

...about Professor Cepko's recent work on developing GFP-activated proteins, read our recent blog post. Check...Multiple Genes to Cells for Gene Therapy 28:40-35:10 : GFP Activatable Tools Being Developed in the Cepko Lab...    References 1.Tang, Jonathan C.Y., et al. “A nanobody-based system using fluorescent proteins as scaffolds...C.Y., et al. “Cell type-specific manipulation with GFP-dependent Cre recombinase.” Nat. Neurosci. 18(9) ...on the Addgene Blog Read our Recent Blog Post on GFP Activable Plasmid Tools Being Developed in the Cepko...

...capture module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph of H1.8-GFP nucleosomes collected...SpyTag-SpyCatcher linkers and spacer proteins ending with a GFP nanobody (Figure 1). This enrichment streamlines the ...In MagIC-cryo-EM, target proteins (expressed as a GFP fusion in this proof-of-concept study) are captured...) 3D model of the obtained structure of the H1.8-GFP-bound nucleosome. Images adapted from Arimura et ...spacers could enable many future variations using nanobodies or scFvs for other targets or could be used for...

...plasmids for: Common antigens such as GFP or mCherry Monoclonals, Nanobodies, Sybodies, or ScFVs Secondary antibodies..., including monoclonal antibodies, scFvs, and nanobodies. Plasmid...scFvs and single-domain antibodies (also known as nanobodies). For example, scientists often use plasmids ...Sybody Generation Toolbox for making synthetic nanobodies (sybodies) against membrane protein targets. ...Elife. 2018 May 24;7. and Generation of synthetic nanobodies against delicate proteins. Nat Protoc. 2020 Apr...plasmids that feature expression of antibodies, nanobodies or ScFvs. Use the search bar or sort buttons ...peptide, from the popular SunTag system, fused to sfGFP for imaging. Learn more about antibodies and their...

...199419 Anti-GFP [N86/38R-2b] GFP Aequorea victoria Mouse IgG2b 199420 Anti-GFP [N86/8R-2b] GFP Aequorea ...206719 Anti-GFP [N86/38R-1] GFP Aequorea victoria Mouse IgG2a 206720 Anti-GFP [N86/8R-1] GFP Aequorea victoria...Rat Mouse 206743 GFP scFv [N86/44] N86/44 scFv GFP Aequorea victoria Mouse 206744 GFP scFv [N86/20] N86... anti-Homer1 nanobody HC87 Homer1 Mouse Llama 135224 GC52 pEGFPN1 anti-Gephyrin nanobody GC52 Gephyrin...1 nanobody AC27 AMIGO-1 Mouse Llama 135227 IC65 pEGFPN1 anti-IRSp53 nanobody IC65 IRSp53 Mouse Llama 135228...SS80 pEGFPN1 anti-SAPAP2 nanobody SS80 SAPAP2 Mouse Llama 145818 SS1 pComb3xss anti-SAPAP2 nanobody SS1...short and long Rat Mouse IgG2a 114492 Anti-GFP [N86/38.1R] GFP Aequorea victoria Mouse IgG2a 114493 Anti-NGL...

... state-of-the-art NIR FPs can be used similar to GFP-like probes by supplying a single gene to cells. ...crosstalk-free three-color imaging with green and red GFP-like FPs (Shcherbakova et al., 2016), as well as ... a scaffold for antigen-dependent fluorescent nanobodies. Nat Methods submitted (2021). Tchekanda, E.,...for miRFPnanos, 35 kDa for miRFPs, and 27 kDa for EGFP) and a possibility to be used as internal tags due... normalization to fluorescence of co-transfected EGFP. e Originally reported as a monomer16, IFP2.0 was...

...available from Andrea Pauli's lab. Neutralizing nanobodies bind SARS-CoV-2 spike RBD and block interaction...Jul 13. (Link opens in a new window) SARS-CoV-2 Nanobodies now available from Ray Owens' lab. Development...libraries . FlipGFP-based activity reporter of SARS-CoV-2 main protease Mpro. (Unpublished) A FlipGFP-based ...

...Cre-dependent expression of diphtheria toxin receptor (DTR)–GFP fusion protein. Azim et al. Nature. 2014 Apr 17. ..., including monoclonal antibodies, scFvs, and nanobodies. New and Noteworthy: Browse our growing collection...