...LanthaScreen) (Degorce et al., 2009), measuring intracellular calcium (Miyawaki, Griesbeck, Heim, & Tsien,...Biarsenical labeling reagents If you want to label intracellular proteins covalently with FRET acceptors, then... nonfluorescent until they bind a 6-residue tetracysteine tag (sequence: CCPGCC) that can be inserted ...interest, resulting in a C-terminal fusion of a tetracysteine tag. In addition, since these tags are so small...large ion channels involved in skeletal muscle contraction.   References 1. Degorce, François, et al. "HTRF...et al. "Hairpin structure of a biarsenical− tetracysteine motif determined by NMR spectroscopy." Journal...

...reverse tetracycline-controlled transactivator) is normally inactive and cannot bind the tetracycline response... inducible promoters. The positive inducible tetracycline ON (Tet-On) system, a versatile tool developed...response elements (TRE) in a promoter. Tetracycline and its derivatives serve as inducing agents to allow...Addgene.org: Inducible Promoters for Bacteria Tetracycline (Tet) On Expression Systems at Addgene Cre-lox...

...by titration with different amounts of tetracycline. Tetracycline regulation is reversible (when the drug...with more elements of control. Tet promoters Tetracycline-dependent promoters are engineered by placing...constitutively silenced proteins. Addition of tetracycline or one of its analogs will induce these transcriptional...transcriptional level.     Fig. 2 Tetracycline on/off promoters and modes of action Check out...

... by ultracentrifugation (75,000 - 225,000 g for 1.5 – 4 hours at 4 °C). After ultracentrifugation, virus...vectors) you can concentrate your virus by ultracentrifugation of the viral stock, followed by resuspension...packaging cell debris before proceeding to ultracentrifugation to avoid contaminating your transduction ... to concentrate a virus solution without ultracentrifugation, you can reduce the volume of cell culture...

...use the autofluorescence extraction feature. Although autofluorescence extraction is an optional feature...correct for this, compensation has to be applied to “subtract” this spillover and correctly distinguish between...possible.  Spectral flow allows for autofluorescence extraction Cells produce a low level of fluorescence called...cytometers include a tool called “autofluorescence extraction” which allows the user to use the autofluorescence...

...enables single-cell anatomical study and neuronal tracing, aka determining where a neuron’s axons and dendrites...labeling so that a cell’s projections are easy to trace and not lost in a jumble of fluorescent neurons.... But notice in figure 4b how difficult it is to trace individual cell’s axons, particularly the two green... as tools for basic science research (neuronal tracing and morphologic studies), as well as translational.... These capsids are useful tools for neuronal tracing and morphology studies when used as part of a two...

...before bleaching. The amount of information one can extract from a biological sample is wholly dependent on...budget, which determines how long and precise we can track individual molecules. Poor photon budgets are a ... couch cushions for spare change, desperate to extract a few more photons from a sample. Chemical fluorophores...FlAsH, ReAsH) and a short genetically encoded tetracysteine (Cys4) peptide tag could be used to label proteins...fluorophores are making chemical dyes increasingly attractive and accessible to cell biologists—and we are ...

... cause cells and muscles to contract depending on the context. Contraction is critical for processes such...1997. Spaghetti Squash, a regulator of cellular contraction You may think that a gene called "spaghetti squash...actomyosin cytoskeleton that regulates cellular contraction. This network is composed of actin and myosin...motility, development, cytokinesis, and muscle contractions (think skeletal muscle, cardiac muscle, and ...

...records allows you to track a cell line’s age and monitor any changes. Your tracking system could be as ...just a few publications which have since been retracted due to cell line contamination: Epithelial Mesenchymal...colleague. Time and money are wasted, papers are retracted and there is a loss of reputation that can impede... good recordkeeping, you need to have a way of tracking how many passages the cells have undergone and...

... so, the WHO recommends one strategy: testing, tracking, and social distancing. With countries like South...block and 62°C heat block Approach: Viral RNA is extracted and subject to isothermal amplification by primers...directly using saliva without having to perform RNA extraction but that may work only if there is a high enough...water bath and 42°C water bath Approach: RNA is extracted from a sample and subject to isothermal recombinase...detect SARS-CoV-2 in the samples. First, RNA is extracted and subject to reverse transcription. This is ...

...has become very useful in lineage tracing; “Zebrabow” was used to trace the development of the corneal epithelium... also been applied to cell marking and lineage tracing in other systems, such as zebrafish and Drosophila...Zebrabow: multispectral cell labeling for cell tracing and lineage analysis in zebrafish. Pan YA, Freundlich...dev.094631. PubMed. Multiplex cell and lineage tracking with combinatorial labels. Loulier K, Barry R,...

...companies which are reliant on contracts with clients is that, when those contracts fall through or are cancelled...(SOP’s) and every item I tested was constantly tracked with barcodes and spreadsheets. It was great to...primary purpose of the company was to fulfill contracts, my work sometimes felt rushed and the focus always...company; new staff hired in anticipation of the contract suddenly have less work to do than was anticipated...

...feature of AAVs as a new tool for neuroanatomical tracing (11). With the ability to produce neuron specific...showing a very specific phenotype. Previous neural tracing methods used viruses that were potentially toxic...available, AAVs could be much better tools for neural tracing in very specific cell populations. Find Ready-to-Use...finger nucleases, and many others (12). The most attractive potential of AAVs is their ability to be used...et al. "A fourth generation of neuroanatomical tracing techniques: exploiting the offspring of genetic...

... cells is minimal (picogram range). It can be extracted from small bacterial cultures (i.e. 2-3 ml) with...cultures and commercial kits you will adopt for the extraction. Restriction enzyme based approaches can require...and guanidine hydrochloride used during plasmid extraction. Figure 2: Use cloning grade DNA for direct... technique must be larger scale, to allow the extraction of enough plasmid to last for multiple experiments...

...complement our RNA Extraction Without A Kit blog post, we've compiled five methods for extracting DNA without...that cause plasmids to be incompatible. 4. RNA Extraction Without A Kit by Leah Schiesow We added to our...it, wrote these tips for working with RNA and extracting RNA without a kit. 3. SARS-CoV-2/COVID-19 Detection...

...cells. They bind to proteins on the surface of extracellular pathogens such as parasites or microbes, or ...from the animal is drawn and the antibodies are extracted. These antibodies recognize a variety of epitopes...Hybridomas are formed by fusing B cells that have been extracted from animals injected with protein antigen to ...presence of protein within a sample, you’ll want to extract your protein of interest from the mixture. Antibodies...permeabilized to allow antibodies to bind to intracellular proteins. The fluorescence signal is detected...

... deadline again. Organize you experiments, keep track of upcoming grant deadlines, and sync everything...Though electronic lab notebooks (ELNs) have gained traction with pharmaceutical companies, academic labs have...allow you to control who can edit the documents and track the revision history. Now you’ll be able to make... knowing the information is backed up and edit tracking will help you identify when others have added ...

...bound to the intracellular surface of the GPCR. Binding of a ligand to the extracellular surface of the...environmental conditions, cells must be able to detect extracellular stimuli. One way they do so is through G protein-coupled...Slocum ST, Che T, Gavin AC, McCorvy JD, Roth BL, Strachan RT (2020) TRUPATH, an open-source biosensor platform...

...the intensity of fluorescence signal and enables tracking of single molecules within living cells without...thereby creating a single-molecule reporter of intracellular processes. Multimerization via the SunTag system....3147. Bioluminescent pH sensor Understanding intracellular pH regulation is important as pH regulation ...to the cell cycle. Good tools for measuring intracellular pH are required in order to determine the relationships.../Cas9 in genome-engineering. One particularly attractive application involves using dCas9-activator fusion... compartment (e.g. the plasma membrane, the extracellular space, or the lysosomes) can take many paths...synchronous release of the reporters which can then be tracked along the secretory pathway (Figure B). The RUSH...

... calculate the amount of bound ligand (after subtracting out non-specific binding). Historically, glutathione-S-transferase...subsequently purify the fusion protein from a bacterial extract. The MBP tag can be removed using Xa protease. ...successfully used this plasmid to biotinylate the extracellular domain of the erythropoietin receptor and subsequently...biotin (50X) because 5 mM is barely soluble at RT. Track expression and purification of biotinylated protein...