...GABA-ergic interneurons. These include mDlx-GFP, hDlx-FLEX-GFP, hDlx-Flex-dTomato, mDlx-GCaMP6f, hDlx-GiDREADD-dTomato...fluorescent tools for labeling structures and compartments in mammalian cells By Angela Abitua Genetically...are often used to label  specific structures, compartments, or specific localization of biomolecules within...usually much easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed...pMAK463 (NBαMouse-IgK, Addgene #140701) pMAK464 (NBαEGFP, Addgene #140702) The pMAK461-64 plasmids were...introduced into the hPSC genome using AAV1 and contain an EGFP reporter. Expression of the dCas9-KRAB activator...

...two photons of red light are needed to excite GFP compared to only one of blue. Only at the focal point...red light instead of blue to excite a molecule of GFP. Because red light has approximately half the energy...infinitely small point of light – think of a single GFP molecule – will appear as a fuzzy 200-300 nm spot...

...mouse brains expressing green fluorescent protein (GFP) (6).   The Dodt and Frank Bradke laboratories later...subpopulation of neurons are brightly labeled with GFP (e.g. Thy1-YFP-H mice).  However, we recently developed... nerve afferents labelled with a AAV8 expressing eGFP under the human ubiquitin C promoter. Anatomical...labeling in whole tissue (15).  CLARITY is also compatible with LSFM, a method termed COLM (clarity-optimized...spatiotemporal control over labeling methods as compared to transgenic animals (17).  These viral plasmids...

...protease protection. Thus far Plasmids 101 has covered GFP and its related fluorescent proteins, which are sometimes... I Use? Plasmids 101: Green Fluorescent Protein (GFP) Fluorescent Protein Guide Expression Vectors from...into a translated protein. As depicted in the accompanying cartoon, they have a multitude of uses including...ProtA IgG-binding domains. TAP has since come to encompass several other tag combinations, though frequently...

... N. C. A., et al. "Evaluation and comparison of the GUS, LUC and GFP reporter system for gene expression...tMAS::rLUC-I::pMAS. Fluorescent proteins (FP, e.g. GFP) can be used in many ways. Often a FP is fused to...

...Figure 5: A) Representative confocal images showing GFP expression from ssAAV5s with hIBA1 or hIBA1a promoters...Quantifications showing high microglia specificity of GFP expression for the indicated ssAAV5s. C) Quantifications...-multi (Walton et al. 2024). This platform is compatible with enrichment, single-cell sequencing, and ...CRISPRi screen to identify additional host factors impacting prime editing and found the protein La (a small...scAAV). However, scAAV vectors have limited size compared to single-strand AAV (ssAAV). To maintain the ...

...P2A-linked accessory genes gvpNJKFGWV marked with Emerald GFP. These plasmids can be used directly in transient...gating for the top quartile (or higher) of BFP and GFP producing cells. After FACS, return the cells to ...structural protein GvpA, which seemed overexpressed compared to accessory proteins. Excess gvpA plasmid added...

...biosensors as an example. Many calcium biosensors include GFP fused to calmodulin (CaM) and the chicken myosin ...biosensors can be targeted to a particular cellular compartment such as the nucleus, mitochondria, endoplasmic... biosensor: which fluorescent protein is most compatible with your lab and experimental setup? Consider...

...blot showing GFP expression in cells that have been transfected with a plasmid encoding GFP. Actin, a common...production strategy has advantages and disadvantages compared to each other. Polyclonal antibodies These antibodies...stability. Additionally, the choice of vector can impact the expression level of an antibody (Ayyar et al...Single-chain variable fragment (ScFv) Figure 2: Comparison between the IgG antibody and scFv. Because... their lower affinity for their target protein compared to whole antibodies (Crivianu-Gaita 2016). Get...Variables (scFvs). Nanobodies Figure 3: Comparison of the Hcab and nanobody. Nanobodies are the...fluorescence. Western blots are frequently used to compare relative levels of protein expression between cell...

...fluorescent protein (GFP), and nLightR uses a red fluorescent protein, mApple. Kagiampaki et al. found nLight...norepinephrine indicators: nLightG and nLightR (Kagiampaki et al., 2023).     Figure 1: Properties...μm (neurons). Figure and caption adapted from Kagiampaki et al., 2023. nLight nLight sensors are based... https://doi.org/10.1016/j.neuron.2019.02.037 Kagiampaki, Z., Rohner, V., Kiss, C., Curreli, S., Dieter...

...highlighted back in 2015, where sdAbs were used to create GFP scaffolds for transcriptional activation.  sdAbs ...to revisit some of the ways these animals have impacted humans through biomedical research. The Camelid...Arbabi-Ghahroudi, 2017)!      Figure 1: Comparison of a heavy-chain only antibody and single-domain...blog over the years and have described how they compare to other antibodies and affinity reagents. But ...And I suspect we will only continue to see their impact grow as new sdAbs become easier to generate and...

...containing the gene of interest (pTet-IRES-EGFP, pPRIME-Tet-GFP-FF3) and one with either tTA or rtTA (FUW-M2rtTA...viral titre to the number of cells transduced by a GFP encoded virus, and the reverse transcriptase activity... as it shows rapid gene expression kinetics as compared to the Tet-off system. One drawback of the Tet...

... (FLAG, Myc, etc.) and fluorescent protein tags (GFP, mCherry, etc.). Fluorescent tags are primarily used...work best attached to, and the species they are compatible with are all reviewed here. What are affinity...

..., hygromycin) and fluorescent protein reporters (GFP, BFP, RFP, no FP). The vectors each contain two SfiI...efficiencies of stable gene transfer using SB100x were comparable to stable transduction efficiencies of integrating...

...or smaller. Some examples include a reporter like GFP, some smaller CRISPR-Cas proteins such as SaCas9,...secondary structure and high GC content, but some companies now offer Sanger sequencing conditions which can...

...endocytosis of a fluorescent dye and transduction of GFP-expressing AAV particles we did not observe any difference...demonstrate that these results were, in fact, due to impaired endocytosis. In parallel assays designed to test...

...strain transformed with pBLADE expressing superfolder GFP instead of mCherry under the PBAD promoter. They ...What advantages does BLADE have over AraC? When compared to the original AraC system, BLADE is simply more...

... University with David Goldstein’s lab, will use GFP-ATP1A3, FLAG-ATP1A3, and ATP1B3 plasmids to explore...paralysis (hemiplegia) that are often painful and accompanied by migraines. Those attacks can be brought on...

...destination vector contains GFP, enabling you to select cells with high GFP expression. These cells have...Oligonucleotides specifying the gRNA seed sequence should be compatible with type IIS enzyme BfuAI. Gateway cloning is...supply it with another plasmid.   Figure 3: Comparison of Multiplex Strategies including Standard PolIII-gRNA...Cas9-containing binary vectors. The system is compatible with both monocot and dicot plants. Liu Lab Golden...

...collection is now complete! pAAV-CAG-GFP (plasmid 37825) and pAAV-hSyn-EGFP (plasmid 50465) are now available...our entire AAV inventory. Our new AAVs include: EGFP-expressing AAV for serotype testing Calcium sensors... testing AAV, which are small (20 ul) samples of EGFP-expressing AAV packaged in various serotypes. This...recently released these controls: pAAV-hSyn-DIO-EGFP (50457-AAV5): The Synapsin promoter directs broad...broad, neuronal expression. AAV pCAG-FLEX-EGFP-WPRE (51502-AAV5): The CAG promoter directs ubiquitous expression...interneurons. pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP (112677-AAV5 and 112677-AAVrg): The EF1a promoter...mCherry in the absence of Cre, and expresses nuclear EGFP in the presence of Cre. Biosensor AAV (calcium ...