...newly developed calcium sensors. Selecting a calcium sensor Genetically encoded calcium indicators (GECIs...unusually fast calcium dynamics, whereas non-neuronal systems tend to have slower, but larger, calcium changes... to use a calcium sensor that is most sensitive to your expected range. For example, calcium in the ER...would want a low affinity calcium biosensor, so that the (1) binding of calcium, (2) allosteric rearrangements...FRET-based calcium reporter derived from troponin C. It is engineered with a reduced number of calcium binding...’ve got a live one (calcium sensors for in vivo studies) There are several calcium reporters specially...Calcium is arguably one of the most important metal ions in biology (sodium and potassium are up there...

... ratios right, you need to do a few calculations. This calculator can do the math for you and it’s simple... download the PEI calculator for AAV packaging Use the PEI transfection calculator for AAV packaging ...spreadsheet will calculate the following μg of each plasmid needed per dish or stack. Calculations are based...corresponding column in the calculator spreadsheet: We hope that this calculator will make it easier to plan...for AAV packaging, considering using this PEI Calculator. AAV packaging typically requires transfecting...whole experiment And here’s the equation the calculator uses to give you the μl needed of each plasmid...experiments. Let us know what you think about the calculator in the comments section below. References 1....

...reporter for calcineurin activity in yeast Article contributed by Leah Schwiesow Calcineurin is a protein...intracellular calcium levels within the cell. When levels of Ca2+ are high in the cell, calcineurin dephosphorylates...attractive drug target. Calcineurin reporters allow scientists to measure calcineurin dynamics and can serve...identify new antifungals that target calcineurin. Current calcineurin gene reporters are based on the bacterial...intermediate (d), and sparse (e) labeling of the calcium indicator, GCaMP6f, in the Drosophila optic lobe...dephosphorylates the protein CZI, which in turn binds to calcineurin-dependent response elements (CDREs) and drives...drives gene expression. Because calcineurin is required for virulence in pathogenic fungi, it is an attractive...

...transfer (FRET)-based calcium biosensors. Perhaps most notably, the Twitch calcium sensors, which are available...-affinity for calcium binding and developed a version of the protein with as few calcium-binding sites...been working on genetically encoded indicators of calcium and other small molecules since the very beginnings...engineered sensors were designed to replace synthetic calcium dyes, which had been in use since the 1980s. “Synthetic...they shift from "off" to "on" in the presence of calcium. This color change can be observed under a microscope...Griesbeck and his colleagues began by isolating a calcium-binding protein, troponin C (TnC), from muscle ...Ratiometric imaging for beginners Griesbeck says his calcium indicators can be used in mouse, Drosophila, C....

...ratio calculators Lab math calculators These calculators can help you with common calculations performed...lab. Molarity calculator Promega's Biomath Calculators Primer resuspension calculator (sign-in required... Cloning ligation reaction calculators These links can help you calculate how much insert and backbone...required.)  DailyCalcs Science Calculator app for iPhone Reagent and Data Repositories These links can help...assembly, or In-Fusion cloning.  NEB Ligation Calculator: This tool works for any cloning method. You ...

...ions, calcium is by far one of the most popular, with dozens of sensors developed for it. Calcium triggers...the presence of calcium. This includes FRET-based sensors such as Twitch. While calcium is king in the ...The calcium is strong here….I can sense it….sound like a plot to an biology scifi movie? Or maybe it’...such as norepinephrine, or an electrolyte such as calcium. This varied component, which we will refer to ...triggers muscle contraction, and calcium channels are activated for neuronal action potentials; it’s an ion... disciplines! Most of the sensors designed for calcium generate a fluorescent output, which will turn ... on the biology you are studying. For example, calcium dynamics in the cytosol and sarcoplasmic reticulum...

... of a given ion - we’ll use calcium biosensors as an example. Many calcium biosensors include GFP fused...region RS20. When calcium is absent, the construct does not fluoresce; when present, calcium induces CaM/RS20...like arabinose or hydrogen peroxide or ions like calcium and zinc. Some biosensors contain a single fluorescent... biosensors from Robert Campbell’s lab display calcium-dependent increases in fluorescence of up to 11,000...common class of biosensors. Blue flame “cameleon” calcium sensors created by Roger Tsien’s lab contain two...proteins (a FRET pair) separated by CaM and M13. When calcium binds, it triggers CaM/M13 interaction that increases...Examples include blue flame mitochondria-targeted calcium sensor CMV-mito-GEM-GECO1  and blue flame extracellular...

...pAAV-hSyn-mCherry Read on to learn more!  Calcium sensor AAV The GFP-based calcium sensors are single-wavelength ...sensors with intensities that change in response to calcium binding. There are a few versions of these jGCaMP7...brightest resting fluorescence of the GCaMP7 family of calcium sensors and can be used for imaging of small neuronal...variant of jGCaMP7 in AAV1 at Addgene! See all calcium sensor AAV at Addgene. GABA sensor AAV GABA is ... directly imaging GABA have been limited. Like calcium sensors, different iGABASnFR have been developed...pAAV-EF1a-mCherry-IRES-Flpo 55637  AAV1  pAAV-EF1a-Flpo Calcium sensors Plasmid Serotype Name 104496  AAV1...

... Biosensor AAV (calcium sensors and GABA sensors) Calcium sensors The latest calcium sensors from Janelia...include: EGFP-expressing AAV for serotype testing Calcium sensors and GABA sensors Optogenetics Recombinases...for monitoring neuronal activity in the form of calcium dynamics. (For more details on this technology,...technology, see our blog post Overview of AAV Encoded Calcium Sensors). We are expanding the inventory of these...pAAV EF1a Nuc-flox(mCherry)-EGFP (112677-AAV5) Calcium Sensors pGP-AAV-syn-jGCaMP7b-WPRE (104489-AAVrg...

... amount of alcohol and decrease the amount of SDS, with a decreased transfer time. Alcohol will slow down...semi-dry transfers, you can adjust the amounts of alcohol and SDS in your transfer buffers. To slow down ...the amount of SDS while decreasing the amount of alcohol. The increased SDS will increase the negative charge...readily with the current.  Pro tip! You need both alcohol and SDS for your proteins to migrate…so don’t try...in an assay, meaning that your normalization calculations will not be correct, and you’ll need to adjust... Increase voltage Increase SDS/decrease alcohol in transfer buffer Small proteins: Over-transferring...Decrease transfer time Decrease SDS/increase alcohol in transfer buffer Mixed proteins: Transferring...

...Plasmid/Kit Alcohol-regulated AlcA promoter AlcR pGGA008 (AlcA) and pGGC011 (AlcR) from red flame...chemically induced promoters include positive inducible alcohol and steroid regulated promoters commonly used in...

...Debate of ’18, Blue Flame depositor John Doench calculated the time to make a plasmid. Doench’s lab in the...long does it take in total? While much harder to calculate, in both our 2018 and 2023 poll, most people said...much does it cost to make a plasmid? Doench also calculated the cost of consumable reagents used to make ...up 19.8% in the same time period. For ease of calculations*, let’s split the difference and put scientific...much do repositories help?   Designing precise calculations for any of the numbers in this post is a mathematical...large-scale savings. How large? Using Doench’s calculations and the 2018 numbers for consumable costs, our...

... of enzyme present in the well and is used to calculate the amount of your protein of interest that is...is present. Using a Standard Curve To do that calculation, you’ll need to include a standard curve in your...samples can then be plotted along this line to calculate the protein concentration. Image from Jagarlamudi...an asymptote, you won’t be able to accurately calculate the protein concentration of any experimental ...sure to incorporate the dilution factor in your calculation from OD to protein concentration if that’s the...linear portion of your standard curve, you can calculate the concentration of protein in the experimental...

... the experiment and use the standard curve to calculate its experimental value. Though it varies from ...enzymatic assays are sigmoidal. Once the trendline is calculated, you can extrapolate the concentration of the...measurements and the trendline y = 0.6561x + 0.019 calculated. The concentration of an unknown sample is determined...standard curve scientists use graphing software to calculate the data’s coefficient of determination, R2. R2...duplicate or triplicate. Replicates allows you to calculate the curve’s coefficient of variation, or %CV. ...mean of all the replicates multiplied by 100. To calculate the %CV of the standard curve, one averages the...

...Acinetobacter spp., Aeromonas spp., Agrobacterium spp., Alcaligenes spp., Aliivibrio salmonicida, Anabaena spp., ...Gram-negative and Gram-positive  Acinetobacter calcoaceticus, Actinobacillus pleuropneumoniae, Actinomyces...Aeromonas hydrophila, Agrobacterium tumefaciens, Alcaligenes eutrophus, Azotobacter vinelandii, Brevibacterium...pSa, pR388 Gram-negative  Acinetobacter calcoaceticus, Aeromonas liquefaciens, A. salmonicida, Agrobacterium...Agrobacterium tumefaciens, A. rhizogenes, Alcaligenes eutrophus, Enterobacter sp., Erwinia amylovora, E... pBBR1 based pBBR1  Gram-negative  Alcaligenes eutrophus, Bartonella bacilliformis, Bordetella...

...doi: 10.1101/2022.08.13.503850.  iGECInano NIR calcium reporter Near-infrared (NIR) fluorescent proteins...’s lab has generated a new genetically encoded calcium indicator using small monomeric NIR fluorescent...photochemical characteristics than previous NIR calcium reporters iGECI and NIR-GECO2G, and its small size...Characterization of a Small Near-Infrared Fluorescent Calcium Indicator. Front Cell Dev Biol. 2022 Jun 29;10:...response size and reduced sensitivity to pH and calcium compared to previous lactate biosensors, and its...additional AAV serotypes of the most recent jGCaMP8 calcium sensors: pAAV-hSynapsin1-axon-jGCaMP8s-P2A-mRuby3...

...luciferase Many intracellular sensors of events like calcium release have been made using fluorescent resonance...generally consist of a responsive protein (e.g. calcium binding protein) fused to two fluorescent proteins...proteins. When the responsive protein binds to calcium it changes conformation and brings the two fluorescent... due to its bright luminescence. Intracellular Calcium Sensor Yang, J. et al. (7) wished to trigger intracellular...this optogenetic tool, they needed to create a calcium sensor with the following characteristics: No ...CalFlux VNT responds to changes in intracellular calcium and is useful for direct imaging of events in cells...

...pAAV-hSyn-DIO-EGFP   Biosensor AAV Calcium sensors Tracking calcium dynamics that govern such activities...studying neural networks. We have expanded our calcium sensor collection to include our first Flp-dependent...Several new options are also available for the calcium sensor jGCaMP7 from Janelia, including three jGCaMP7...technology, see our blog post Overview of AAV Encoded Calcium Sensors). Plasmid Serotype Name 104489 AAVrg...

...tap water. Add ethyl alcohol or acetone drop by drop onto the smear until the alcohol runs clear and immediately... one type, Streptococcus pneumoniae, while the alcohol used in the staining process bleached another, ...peptidoglycan layer, staining the cell purple. When the alcohol is added, it dehydrates the peptidoglycan layer...positive bacteria. In a gram negative bacteria, the alcohol destroys the outer lipid membrane, and the thin...

...and iGECI calcium indicator (Shemetov et al., 2021). In addition to FRET, intensiometric calcium indicators...performance, and applications of near-infrared calcium indicators in this recent review (Shcherbakova ... genetically encoded near-infrared fluorescent calcium ion indicator. Nat Methods 16, 171-174 (2019). ....A. et al. A near-infrared genetically encoded calcium indicator for in vivo imaging. Nat Biotechnol 39...