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We narrowed to 134 results for: Green Fluorescent Protein

Showing: 101 - 120 of 134 results
  1. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

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    Blog Post
    ...make. For example, large insertions, like a fluorescent protein knock-in, suffer from low efficiency when... your desired insertion feature (e.g., a fluorescent protein), you can amplify the desired region using... of the commonly used cassettes, such as fluorescent proteins (EGFP, mCherry etc.), recombinases (Cre,... Quadros et al. recommend ribonucleoprotein (RNP) complexes of Cas9 protein and the separated guide RNA...cleavage sites (slashes) are shown. In this case, the green guide would be selected, since it cleaves at the...You can also use Addgene plasmids to purify Cas9 protein. The table below contains some of our favorites...alleles using long ssDNA donors and CRISPR ribonucleoproteins.” Genome Biol. 18 (2017) (1):92. DOI: 10.1186...
  2. A Primer on Optogenetics: Introduction and Opsin Delivery

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    Blog Post
    ...co-localization immunofluorescence, cell-type specificity. Viruses that only express the fluorescent proteins are ...Thankfully the Deisseroth constructs coexpress a fluorescent protein, which allows for easy identification of ...vectors for anterograde axonal tracing with fluorescent proteins in nontransgenic and cre driver mice. Current...optogenetics plasmids! Find AAV resources! Find fluorescent protein resources!    ...of an action potential. Halorhodopsins (HR) are green/yellow-light sensitive chloride ion pumps, activation...general rule of thumb, if you can detect a strong fluorescent signal (without antibody manipulation), then ...
  3. Using AAV for Neuronal Tracing

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    Blog Post
    ...that express one of a few different fluorescent proteins (red, green, blue). If a researcher injects a ...and neuron-specific transgene expression. Fluorescent proteins are widely used for tracing and their expression...Maday et al., 2014). It is currently unclear which proteins facilitate transport of neuronal tracing compounds...either produce a colorimetric product or are fluorescent. While dyes usually spread unidirectionally, ...biotinylated dextran amines (Veenman et al., 1992) or fluorescent molecules like rhodamine-isothiocyanate (RITC... For predominantly retrograde tracing, albumin protein labelled with horseradish peroxidase (HRP; Kristensson...germ agglutinin WGA, Schwab et al., 1978) or fluorescent molecules such as Fluoro-Gold (Schmued and Fallon...
  4. Antibodies 101: The Four ELISAs and When to Use Them

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    ... expresses the endogenous protein, since folding of the recombinant protein may differ from the native... have since adapted it for a host of different proteins and small molecules from a variety of sample types...peroxidase (HRP) or alkaline phosphatase (AP). Fluorescent tags may also be used, but for simplicity we ...multiwell plate is coated with a competing antigen (green).  The sample (purple) is preincubated with a reporter...antibodies and thus are not suitable for low abundance proteins. Because the method uses a single antibody, it... and a series of wash steps removes non-target proteins and molecules from the assay. The user then adds...endogenous sample of wild type cells, purified protein, or a peptide. The positive control will confirm...
  5. Fluorescent Proteins: FRET

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    ...to report on protein binding or conformational changes. Organic dyes, fluorescent proteins, and other molecules...is often used to study (1) protein-protein interactions where each protein is separately fused to a donor...brilliant and fast-maturing red fluorescent protein Variety of fusion proteins, positive controls for many... Plasmid Collections Fluorescent Proteins FRET Fluorescent...changes within a protein where the donor and acceptor are both fused to the same protein (also referred ...FRET Pairs This table lists some popular fluorescent proteins used as FRET pairs along with their properties...orientation of the fluorophores through linkers, fluorescent protein expression level and maturation time, and...
  6. Our Most Popular Blog Posts from 2019

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    Blog Post
    ...graphics. PS: they’re free. 3. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection...Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi by Jennifer Tsang Learn about mNeonGreen, mTFP1..., optogenetics, chemogenetics, plasmids, fluorescent proteins, careers, and more. These resources can ...Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins by Cary Valley, Shreya...mWasabi and find plasmids containing these fluorescent proteins at Addgene. 7. Designing Your Chalk Talk...strategies for generating GFP fusion proteins. From considering protein size and shape, linker sequences,...chalk talk, and tips for practicing.  6. The Fluorescent Vegetables in Aptamer Soup by Aliyah Weinstein...
  7. Course-Based Undergraduate Research Experiences (CUREs)

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    ... of a fusion protein: Histidine tag, malate dehydrogenase, and green fluorescent protein. Biochemistry... graduate school. As a stable, easily purified protein with easily measured activity, MDH is particularly...structure, function, and interaction with other proteins.   It is important to note that research ...
  8. Plant Plasmids and Resources

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    ...Resources Visit our Fluorescent Protein Guide for more plasmids containing fluorescent proteins. A set of fluorescent... expression, protein localization, proteinprotein interactions, and other physiological processes in ...analysis of protein-protein interactions in planta via ratiometric Bimolecular Fluorescence Complementation... bimolecular fluorescence complementation (MoBiFC) toolkit for analyzing protein-protein interactions ...reporter plasmids that feature fluorescent, luminescent, and color-producing proteins for detection of gene expression...plants. ID Plasmid Description PI Kits for ProteinProtein Interactions The following plasmid kits contain...variety of reporter plasmids for the study of proteinprotein interactions in plants. ID Kit Description ...
  9. Hot Biosensors 2022: Year-End Roundup

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    Blog Post
    ... blog Fluorescent Proteins 101: Fluorescent Biosensors Resources on Addgene.org Fluorescent protein...short), where a fluorescent protein is inserted into one of the loop regions of any G-protein-coupled receptor...fusing two bright fluorescent proteins with different thermal quenching rates (mNeonGreen and tdTomato), ...NIR calcium reporter Near-infrared (NIR) fluorescent proteins promise better performance for in vivo use...calcium indicator using small monomeric NIR fluorescent proteins (miRFP670nano and miRFP720) and the Ca2+...engineering of potassium binding proteins, they developed a family of fluorescent potassium indicators with ...protein plasmids and resources Fluorescent protein guide: Biosensors Collection ...
  10. Hot Plasmids - August 2020

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    ...'ll find in this post: Yeast-based protein evolution Fluorescent tools for mammalian cells Nanobody purification...enables tracking of protease activity with a fluorescent protein--no affinity purification required! The team...cells By Angela Abitua Genetically encoded fluorescent proteins are often used to label  specific structures...biomolecules within cells. However, sometimes these fluorescent proteins might not localize to the right places, ...example, to improve nuclear localization of fluorescent proteins, they found that adding a triple nuclear...Find the sensors at Addgene! New yellow fluorescence protein-based calcium sensors (jYCaMP1), variants...nm and enable two-color imaging with red fluorescence protein-based calcium sensors. These new calcium...
  11. New Optogenetic Tools for Cytoskeleton and Membrane Control

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    ...photoswitchable elements into many other types of proteins. Here, we’ll highlight a few recently-deposited...license. A naturally occurring microtubule-severing protein, Katanin is of course named for the samurai sword...optogenetic module to activate or recruit your protein of interest, maybe you want to inhibit it. Restricting...development and a knockout is not feasible or when the protein of interest supports a variety of processes in ...CIB1 clustering. Fuse either CRY2 or CIB1 to your protein of interest and express the other separately, then...generate CRY2-CIB1 oligomers that trap the fusion protein away from its usual location, preventing it from...such a strategy to dissect the roles of three Rab proteins during a key stage of zebrafish development (Aljiboury...
  12. Brain Initiative Collection

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    ...encoding Cre-dependent expression of the fluorescent protein EYFP from the CAG promoter Viviana Gradinaru...AAV genome that ubiquitously expresses the fluorescent protein eYFP from the CAG promoter Viviana Gradinaru...hSyn1-eYFP An AAV genome that expresses the fluorescent protein eYFP from the hSyn1 promoter Viviana Gradinaru...
  13. Sensing Neuronal Dopamine

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    ...researchers from some labs have developed fluorescent-protein based DA sensors (Figure 2). The dLight1 ...sensors, the new dLight1 sensor is a single fluorescent protein-based sensor. This sensor was engineered ...a GFP variant and a DA receptor such that GFP fluoresces when the receptor is bound by DA. Two variants...affinity and high-magnitude responses (increases in fluorescence) at low concentrations (e.g., 100 nm) of DA....variant bound to a DA receptor, such that the GFP fluoresces upon DA-receptor binding (Sun et al., 2018). ...These sensors also exhibit high magnitudes of fluorescent responses. Studies on the kinetics of these sensors...Sun, Fangmiao, et al. "A genetically encoded fluorescent sensor enables rapid and specific detection of...
  14. Quantifying DNA? Here are Five DNA Quantification Methods to Consider

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    ...DNA Fluorescence dyes Another way to quantify DNA would be to use fluorescent dyes that fluoresce when...and their corresponding fluorescence. That way, you can compare the fluorescence of your sample against...acids (DNA and RNA) absorb maximally at 260 nm. Proteins on the other hand absorb best at 280 nm and organic...sequencing. Unlike absorbance-based methods, fluorescence-based methods require a standard curve, a set...this method doesn’t tell you about contaminating proteins or chaotropic salts. Capillary electrophoresis...These fragments are quantified over time using a fluorescent dye that intercalates into the DNA such that ... using capillary electrophoresis which shows fluorescence units over time. Smaller fragments migrate more...
  15. Technical Design of a Western Blot

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    ...for the amount of protein you are expecting.  Fluorescent proteins Fluorescent proteins, while more expensive...binding to your protein of interest. Protein preparation Before you can blot your proteins, you’ll need ...normalize your protein of interest to overall protein expression for each sample and protein. While ubiquitously...normalize your protein of interest to total protein expression. Total protein expression can be measured using...rely on primary antibodies conjugated to fluorescent proteins, instead of the more common chemiluminescent...consider the subcellular location of your protein. Proteins located in membrane-bound subcompartments,...considered if your protein is difficult to solubilize, such as a membrane-bound protein. There are many ...
  16. Hot Plasmids - May 2022

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    ...GFP-nanobody rather than a specific protein of interest. This fusion protein can then be co-expressed in essentially...stabilizes the TurboID-nanobody protein and allows TurboID to start labeling proteins in close proximity to the...application. Each visual barcode consists of a fluorescent protein coupled to a specific subcellular localization...localization peptides with four different fluorescent proteins, and inserting each visual barcode into ...reporters that drive the expression of the fluorescent protein by a specific transcription response element...approach has been hampered by the need to tag each protein of interest and, for in vivo work, generate a dedicated...zebrafish line expressing a GFP-tagged version of a protein of interest. The binding of the GFP nanobody to...
  17. Deep Dive: qPCR

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    ... be used as a proxy measure for differences in protein expression. (If you're just getting started, check...fluorogenic primers Low Single self-quenching fluorescent molecule connected to a primer  No Yes ...normalize for that. Passive reference dyes are inert fluorescent dyes that are not dependent on amplification ... It is possible to multiplex qPCR when using fluorescent probe-based chemistry - theoretically, up to ... Tip🔥 The ABI machines automatically record fluorescent data on all channels - helpful if you forget ...quantification of relative mRNA abundances using SYBR Green I and real-time RT-PCR. J. Immunol. Methods 283:...
  18. 27 Hot Plasmids from 2016

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    ...method for identifying protein-protein associations. This method uses a bait protein fused to a biotin ligase...PrM N and C   Detecting protein-protein interactions with DULIP Proteins rarely act alone. Molecular...which “bait” proteins are tagged with Protein A-Renilla luciferase and the “prey” proteins are tagged with...with Rosella, a pH-sensitive fluorescent biosensor A pH-sensitive fluorescent biosensor-based assay system...ligase to biotinylate proximate proteins in the cell. These candidate proteins can be enriched via biotin ...of fusion proteins, requires less biotin, and shows enhanced labeling of proximate proteins. Overall BioID2...BioID2 improves the efficiency of screening for proteinprotein interactions. The Roux lab has deposited HA...
  19. Antibodies 101: Secondary Antibodies

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    ...molecule, the antibody binds to a protein, and then voila! Wherever your protein of interest is, a signal will...signaling molecules are represented in light green; and proteins are represented in orange. Created with BioRender.com...molecules to identify each protein of interest, this gel-based method identifies proteins solely by weight and... isn’t giving you enough signal to detect your protein of interest, it may be time to look at the indirect...antibodies. While a primary antibody binds to the protein of interest, a secondary antibody binds to the ...signaling molecules associated, indirectly, with each protein of interest.    Figure 1: The direct ...three, secondary antibodies are indicated when the protein of interest is found in low amounts or if the conjugated...
  20. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

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    Blog Post
    ...sequences for several different fluorescent proteins. Visualizing fluorescence can be challenging in plants...because of the auto-fluorescence of chlorophyll, so we looked at newer fluorescent proteins that get around...construct on the same piece of DNA. We do a lot of protein-protein interaction work and we’ve gotten some tools...Reading: Quick, Versatile Plant Transgenesis with GreenGate Plasmids  ...
Showing: 101 - 120 of 134 results