We narrowed to 740 results for: NSI

...retain fluorescent intensity during exposure to light, is another factor to consider. If your application...interiors to many inquisitive scientists. Seeing inside these specimens has offered illumination on biochemical...will outline some fundamental characteristics to consider when selecting a dye or ligand to make your selection...proteins are SNAP-tag® and HaloTag®. These tags consist of a genetically encoded enzyme variant that covalently... is bound through green light emission. Key considerations in selecting fluorescent dyes/ligands Effects...-conjugation affects protein function and its propensity to bind cellular components. Some dyes target...stably combined with nanoparticles, maintaining intensity at ~85% of its original value after 60 minutes...

...subcloning, expansion, and confirmation. This procedure can take several months but should be considered in labs...benefits and drawbacks of different methods (for comprehensive protocols for all of the methods discussed here...results in less than a day, but tends to be less expensive than an ELISA kit. In this approach, viral RNA...lentiviruses in parallel, this approach should be considered. Similar to the p24 assay, titering via measurement...When using qPCR based methods for viral titration consider using a reverse transcriptase mutant in a separate...carryover. Functional titer Functional titer is considered a more accurate method to quantify virus as it...of this approach is that it can be quite labor intensive. Target cells are transduced with serial dilutions...

...the least expensive of all the options discussed as it only requires PCR and the inexpensive DpnI enzyme...here to help with that! Four important factors to consider for any experiment are: Speed Cost Difficulty...For cloning in particular, another important consideration is: The number of inserts ...as some methods...the following cloning methodologies have been extensively described in other articles; so if you are interested...little as 30 minutes! Alternatively, you might consider Gibson cloning. In this technique, you first perform...order determined by their homology. You might also consider FastCloning, first described by Chaokun Li and...Finally, TOPO cloning is another good option to consider when time is of the essence. This method requires...

...cellular-level events and behavior. It’s a problem that he considers “one of the greatest challenges of neuroscience...particular advantage for observing events taking place inside cells that are on the move, such as T lymphocytes... calcium-binding sites as possible for added sensitivity. As reported in Nature Methods, the FRET changes...essentially all of the standard organisms for which extensive genetic tools are available. While he recognizes... he said. “It gives you more information than intensity-based readouts.” Griesbeck also points out that...approach he and his colleagues took to optimize sensitivities can now be applied to dozens of other published...for visualizing the transcription factor cAMP-responsive element-binding protein (CREB) in living cells...

...classes of optogenetic tools, and what factors to consider when choosing one.     Figure 1: Example...or inhibit specific neurons without needing to consider secondary signaling pathways. There are many different...different opsin (natural and engineered proteins responsive to light in combination with retinal) variations...reviewed in Abreu & Levitz, 2020), including light-sensitive chimeric GPCRS (OptoXRs) (Tichy et al., 2019),... the extracellular domain of endogenous light-sensitive receptors, like mammalian rhodopsin, and the intracellular...Created with BioRender.com.   Important considerations for using optogenetic tools Once you’ve selected...specific tool. There are many, many options to consider, and the following factors will help you determine...

...your plasmid yield is sub-optimal? If you are consistently getting sub-optimal plasmid yields from your...Addgene uses and recommends DH5𝝰! This strain consistently gives good, high quality preps due to its endA1... this handy guide from OpenWetWare for a more extensive listing of common E. coli strains. Increase the...media or adding supplements can improve your cell density, leading to an improved plasmid DNA yield. Culture...supplements to the media You can achieve higher cell density using a nutrient-rich broth such as Terrific Broth...in moderation) may also serve to increase cell density and plasmid yield. Start with a single, fresh colony...Increasing the incubation time can improve the cell density and therefore the plasmid yield. You can optimize...

... One consists of NLuc fused to enhanced green fluorescent protein (GpNluc) and the other consists of NLuc... (FRET) probes. These probes generally consist of a responsive protein (e.g. calcium binding protein) ...membrane-inserted voltage sensing domain (LOTUS V; Luminescent Optical Tool for Universal Sensing of Voltage). With...how bright you can make a reaction but for how sensitive you can make a reaction. NLuc requires 100-fold...) fused to two fluorescent proteins. When the responsive protein binds to calcium it changes conformation.... In well-designed FRET-based biosensors, the intensity of the FRET signal can be correlated to specific...No photoexcitation High Ion specificity High sensitivity Easy expression Gives ratiometric data The result...

...each event to a channel number based on intensity. Higher intensity events are assigned to higher channels...a user will compare the shifting intensity of events. Since intensity positively correlates with expression...cabinet with cloning rings or following labor-intensive limiting dilution protocols? If this sounds like... the cell’s size, complexity and fluorescence intensity.   What flow cytometers detect Cell size and... a high degree of internal complexity such as extensive membranes produces a greater SSC.  Protein expression...The electrical system converts the fluorescence intensity to a voltage pulse, called an event, and assigns...reporter on the N- or C-terminus of the protein. One downside of fusion proteins is that the tag can alter the...

... to be considered as well. In this post, we’ll touch on antibody storage, buffer considerations, and give...records, and consider the following questions as you move forward:  Was a light-sensitive conjugate on...you’re planning to run with the antibodies time-sensitive, using precious samples, or being compared to ...data in a manner that demands high accuracy and consistency?  If the answer to any of these questions is...saving the poor, abandoned antibody for a…less sensitive moment.   Fig 1: Blugene fading into darkness...fix it. HRP is easy to find, simple to use, not sensitive to light in its inactive state, and cheap. Unless... panel in the machine. Fluorophores are light sensitive, so antibodies conjugated to them need to be stored...

...more sensitive channelrhodopsin actuator for triggering action potentials at low light intensities, the...cumbersome. To overcome many of the limitations intrinsic to DNA-guided systems such as TAL effectors or...antibodies fused to sfGFP. This system amplifies the intensity of fluorescence signal and enables tracking of... H+ specific response cytoplasmic retention insensitivity to other ions In BRET, A mutant Renilla luciferase...dimers by fusing the SsrA peptide with the light-sensitive LOV2 domain of Avena Sativa phototropin 1 (AsLOV2...activator protein to drive transcription. The anchor consists of dCas9 fused to CIB1 and is targeted to specific... a photoswitchable fluorescent protein which transitions from emitting orange (λ= 565nm) to far-red (λ...