...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....

...screening in Plasmodium falciparum.... Plasmid Collections EXPRESs Plasmids Plasmodium falciparum EXPRESs Plasmids...Plasmids for Plasmodium falciparum You may also like... Gavin Wright Lab plasmids This is a collection of plasmids...by the blood stage of the malaria parasite, P. falciparum . Please see the following reference for more...essential for erythrocyte invasion by Plasmodium falciparum. Crosnier C et al., Nature. 2011 Nov 9;480(7378...

...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...

...31. Gordon Fishell Calcium GCaMP-X genetic encoded calcium sensor Improved calcium sensor GCaMP-X overcomes...Xiaodong Liu Calcium GCaMP5 genetically encoded calcium sensor Optimization of a GCaMP calcium indicator ...GCaMP calcium indicators. Nat Methods. 2009 Dec;6(12):875-81. Loren Looger Calcium NEMO calcium sensors...Schreiter Calcium GCEPIA1-SNAP ER for sensing ER calcium A genetically encoded ratiometric calcium sensor...Target Description Article PI Calcium jGCaMP8 Fast Genetically Encoded Calcium Indicators (Constitutive or...Encoded Calcium Indicators. Janelia Research Campus. Online resource. 2020 GENIE Project Calcium jGCaMP7...High-performance GFP-based calcium indicators (Constitutive or Cre-dependent) High-performance calcium sensors for imaging...

...jRGECO1a Calcium Sensor Kim , GENIE 100854 pAAV.Syn.NES-jRGECO1a.WPRE.SV40 Syn jRGECO1a Calcium Sensor ...pGP-AAV-syn-jGCaMP7s-WPRE Syn GCaMP7s Calcium sensor Kim 104489 pGP-AAV-syn-jGCaMP7b-WPRE Syn GCaMP7b Calcium sensor Kim 104491...Cre-dependent Calcium sensor Kim 104493 pGP-AAV-syn-FLEX-jGCaMP7b-WPRE Syn GCaMP7b, Cre-dependent Calcium sensor...Cre-dependent Calcium sensor Kim 104496 pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE CAG GCaMP7f, Cre-dependent Calcium sensor...GCaMP7f Calcium sensor Kim 104492 pGP-AAV-syn-FLEX-jGCaMP7f-WPRE Syn GCaMP7f, Cre-dependent Calcium sensor...GP-AAV-syn-jGCaMP8s-WPRE Syn GCaMP8s Calcium sensor GENIE 162375 pGP-AAV-syn-jGCaMP8m-WPRE Syn GCaMP8m Calcium sensor GENIE 162376...GCaMP8f Calcium sensor GENIE 162377 pGP-AAV-syn-FLEX-jGCaMP8s-WPRE Syn GCaMP8s, Cre-dependent Calcium sensor...

..., including calcium sensors, under the control of various promoters. Sensor Green Calcium Sensors GCaMP8...soma-GCaMP6 Twitch2B Yellow Calcium Sensors jYCaMP1 axon-jYCaMP1 Red Calcium Sensors CaMPARI jRCaMP1 jRGECO1...viral service encoding biosensor tools, including calcium sensors. Viral...jRGECO1 sRGECO Far-Red/NIR Calcium Sensors HaloCaMP1a HaloCaMP1b NIR-GECO Acetylcholine Sensors iAChSnFR..._ACh3.0 Syn GRAB_ACh3.0 none Constitutive 9 Li Calcium Sensor: GCaMP8f 162376 pGP-AAV-syn-jGCaMP8f-WPRE...CAG jGCaMP8f none Constitutive 1, 9, rg* Looger Calcium Sensor: GCaMP8s 162374 pGP-AAV-syn-jGCaMP8s-WPRE...CAG jGCaMP8s none Constitutive 1, 9, rg* Looger Calcium Sensor: GCaMP8m 162375 pGP-AAV-syn-jGCaMP8m-WPRE...

...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...

... beta CALC2, CGRP-II, CGRP2, FLJ30166 CALCR calcitonin receptor CRT, CTR, CTR1 CALCRL calcitonin receptor-like...CD88 CALCA calcitonin-related polypeptide alpha CALC1, CGRP, CGRP-I, CGRP1, CT, KC, MGC126648 CALCB calcitonin-related...S5.7, TAPA1, TSPAN28 CHP calcium binding protein P22 SLC9A1BP CHP2 calcineurin B homologous protein 2 -...suppression of tumorigenicity 2 - STC1 stanniocalcin 1 STC STC2 stanniocalcin 2 STC-2, STCRP TAC1 tachykinin,...SLAMF2, hCD48, mCD48 CHP calcium binding protein P22 SLC9A1BP CHP2 calcineurin B homologous protein 2 -...CMM3, MGC14458, PSK-J3 CHP calcium binding protein P22 SLC9A1BP CHP2 calcineurin B homologous protein 2 -...nuclear factor of activated T-cells, cytoplasmic, calcineurin-dependent 1 MGC138448, NF-ATC, NFAT2, NFATc NFATC2...

... CAG Calcium sensor, Cre-dependent GCaMP Kim , GENIE 104487 pGP-AAV-syn-jGCaMP7s-WPRE Syn Calcium sensor...pGP-AAV-syn-jGCaMP7f-WPRE Syn Calcium sensor GCaMP Kim , GENIE 135632 pAAV-S5E2-GCaMP6f E2 regulatory element Calcium sensor ...pAAV.Syn.NES-jRGECO1a.WPRE.SV40 Syn Calcium sensor RGECO Kim , GENIE 162375 pGP-AAV-syn-jGCaMP8m-WPRE Syn Calcium sensor GCaMP...Biosensors 104491 pGP-AAV-syn-FLEX-jGCaMP7s-WPRE Syn Calcium sensor, Cre-dependent GCaMP Kim , GENIE 104492 ...104492 pGP-AAV-syn-FLEX-jGCaMP7f-WPRE Syn Calcium sensor, Cre-dependent GCaMP Kim , GENIE 104495 pGP-AAV-CAG-FLEX-jGCaMP7s-WPRE...pGP-AAV-CAG-FLEX-jGCaMP7s-WPRE CAG Calcium sensor, Cre-dependent GCaMP Kim , GENIE 104496 pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE...

...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...

...Plasmodium falciparum genome. To put this last point into perspective, the Plasmodium falciparum genome contains...in their work editing the genome of Plasmodium falciparum (the parasite that causes malaria). I attribute...sites, scores and the statistics that go into calculating them. For this, some online tools such as Cas-OFFinder...editing in the human malaria parasite Plasmodium falciparum using the CRISPR-Cas9 system." Nature biotechnology...

...tried to find an app that could help me with calculations. This is how I came across the Gibco Cell Culture...culture and include a counter, lab timer, dilution calculator, and a couple of other nice features. This app...sequence for your enzyme of interest. Lastly, the Tm calculator helps you set up an optimal PCR depending on ...apps: Findings Lab Notebook Cell Culture Split Calculator by DRDP Restriction Enzyme Tool by Promega Material...

...fluorescence protein-based calcium sensors (jYCaMP1), variants of the calcium indicator jGCaMP7, that are...with red fluorescence protein-based calcium sensors. These new calcium sensors include syn-jYCaMP1s (slow...

...Zebrafish receptors Zebrafish antibody plasmids P. falciparum merozoite surface protein plasmids P. vivax merozoite...erythrocyte Babesia microti blood stage proteins P. falciparum sporozoite cell surface and secreted proteins...essential for erythrocyte invasion by Plasmodium falciparum. Crosnier C, Bustamante LY, Bartholdson SJ, Bei...

...SPARC to image the calcium response in the dendrites of T5 neurons by placing the calcium indicator GCaMP6f...intermediate (d), and sparse (e) labeling of the calcium indicator, GCaMP6f, in the Drosophila optic lobe...

.... Factor Xa loves calcium, but metal-chelate matrices, not so much. Maybe a calcium-independent proteinase...This allows washing away of unbound material to calculate the amount of bound ligand (after subtracting ...

...Control AAVs for serotype testing jGCaMP7c for calcium detection PSAM4 GlyR for chemogenetic inhibition...high contrast between its resting state and its calcium-bound state, which was achieved by lowering the...  AAV1  pAAV.hSynapsin.SF-iGluSnFR.S72A New calcium sensors Plasmid Serotype Name 104496  AAV1...

...might not have to do the calculation yourself as many fluorometers will calculate your sample concentration...than 1.5. Then, using the A260 reading, you can calculate the DNA concentration. Generally, A260 of 1.0 ...

... followed by a talk from Professor Karin about alcohol and obesity as driving factors for chronic inflammation...chronic hepatitis caused by hypernutrition and alcohol consumption. Chronic inflammation is a dangerous...cells that are not excitable but responsive to calcium. We’d love to expand our cancer and immunology ...

...robust, with an eNL-based calcium sensor displaying a 500% signal change upon calcium binding. Suzuki et al... to previous publications. They also developed calcium, cAMP, and ATP sensors based on Nano-lantern, showing...

...-mCherry-hPMCA2w/b Expresses a plasma membrane calcium pump hPMCA2w/b Baljit Khakh 112000-AAV1 pAAV-hSynapsin1...135420-AAV1 pAAV-syn-FLEX-jYCaMP1s Yellow protein calcium sensor expressed under human Synapsin1 promoter...pAAV-syn-FLEX-axon-jYCaMP1s Axon-targeted yellow protein calcium sensor expressed under human Synapsin1 promoter...Podgorski 135424-AAV1 pAAV-syn-jYCaMP1s Yellow protein calcium sensor expressed under human Synapsin1 promoter...

...not use CsCl are commercially available. Titer calculation (1 or 10 days) Physical titer (particle count...260nm corresponds to 1.1 x 1012 particles/mL. To calculate the particle number: OD260 reading x dilution ... days later. Plaques are counted and titer is calculated in terms of plaque forming units (pfu), taking...

...used to calculate the concentration of the sample. This concentration can then be used to calculate the viral...Maintaining a high droplet count is important for calculating the concentration of your sample. Even if you...

...Andrew Leifer’s lab utilized the worm to study calcium signaling in the entire adult brain, using a plasmid...plasmid that expresses a genetically encoded calcium sensor, GCaMP6 (Nguyen et al., 2015). If you are ...PMC5349099. Nguyen, Jeffrey P., et al. "Whole-brain calcium imaging with cellular resolution in freely behaving...

...Transfer (BRET) CalFlux VNT : An intracellular calcium sensor created by fusing NanoLuc® and Venus fluorophore...interest can also be inserted into the plasmid. Malcolm Moore 51819 Syn-ATP Firefly (modified) CMV Optical...Lentiviral expression of firefly luciferase Javier Alcudia 113442 pcDNA3.1 NL NanoLuc® CMV Mammalian expression...

...it to create a standard curve on your blot to calculate the amount in your sample. (Do ensure your blot.../Reprobe PVDF membrane Increased time Calculate absolute quantification Running a standard curve...wells; purified protein not always available Calculate relative quantification Normalizing to ubiquitously...

...by the blood stage of the malaria parasite, P. falciparum . Please see the following reference for more...essential for erythrocyte invasion by Plasmodium falciparum. Crosnier C et al., Nature. 2011 Nov 9;480(7378...

... CRISPR-based method for cell lineage tracing Valcyrie is a CRISPR-based marker excision method for marking...target. Read the paper in Development Find the Valcyrie plasmids If you have a new CRISPR tool you’ve...

...15R] QKI-7 Human Mouse IgG2a 177508 Anti-NALCN [N185/7R] NALCN Rat Mouse IgG2a 177509 Anti-Dopamine D2 ...Mouse IgG2b 225412 NALCN (Rattus norvegicus) recombinant monoclonal antibody. NALCN Rat Mouse IgG2b 225413...Alpha1 Human Mouse 206721 NALCN ion channel scFv [N185/7] N185/7 scFv NALCN ion channel Rat Mouse 206722...

... lot of their plastic packaging (from Corning®, Falcon®, or Axygen® product packaging) for free! It has... If you use serological pipettes from Corning®, Falcon®, or Axygen®, set up a bin or box in your tissue...

...by the blood stage of the malaria parasite, P. falciparum . Please see the following reference for more...bio-His 53410 PCYOX1L-bio-His 53411 PF4-bio-His 53412 CALCR-bio-His 53413 PPIB-bio-His 53414 PTGS1-bio-His 53415...

...-encoded reporters of molecular signals; e.g., calcium indicators In this overview, we will focus on the... truncated forms of cytosolic STIM1 to control calcium channels ~488 LOV domains LOV-based Light-oxygen-voltage-sensing...light-controlled molecules. ~450–495 BACCS light-activated calcium channel switch iLID Improved light-inducible dimers...

...manipulation in the lab. This is commonly done using calcium chloride which permeabilizes the cell membrane ...and physical methods in the lab. Chemicals like calcium phosphate and diethylaminoehtyl (DEAE)-dextra neutralize...

...will image these neurons using GCaMP7, measuring calcium signaling related fluorescence during morphine ...enable multiplexed imaging of neurotransmitter, calcium, and membrane voltage,” says Tian. Figure...