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Showing: 21 - 40 of 51 results
  1. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...GABA-ergic interneurons. These include mDlx-GFP, hDlx-FLEX-GFP, hDlx-Flex-dTomato, mDlx-GCaMP6f, hDlx-GiDREADD-dTomato...with plasmids encoding FPs tagged with the plasma membrane targetin sequence derived either from p63(1-29...non-specific labeling. For labeling the plasma membrane, they tested different target sequences for improved...pMAK463 (NBαMouse-IgK, Addgene #140701) pMAK464 (NBαEGFP, Addgene #140702) The pMAK461-64 plasmids were...introduced into the hPSC genome using AAV1 and contain an EGFP reporter. Expression of the dCas9-KRAB activator...
  2. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    Published
    Jan. 19, 2021, 2:15 p.m.
    ...blot showing GFP expression in cells that have been transfected with a plasmid encoding GFP. Actin, a common...SDS-PAGE, and are subsequently transferred to a membrane. A primary antibody recognizing the protein of...is added, and will bind to that protein on the membrane. Then, addition of a secondary antibody allows...bind to proteins on the cell surface, or the cell membrane can be permeabilized to allow antibodies to bind...
  3. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    Published
    Oct. 27, 2015, 3 p.m.
    ...excellent choice for fusions with integral membrane proteins and membrane associated proteins (i.e. GPI-anchored...including the secretory pathway (Figure 2), the inner membrane space of mitochondria and chloroplasts, the extracellular...Lindsey M. Costantini, and Erik L. Snapp. "Superfolder GFP is fluorescent in oxidizing environments when targeted...
  4. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...they cannot cross cell membranes, so they are best used either with plasma membrane proteins (such as GPCRs...His  44 35-63 Plasma membrane protein, permeabilized cells  Cy5NTAc 32 25-45 GFP (fluorescein is similar... Most importantly, these compounds cross cell membranes, thus enabling FRET measurements in intact cells...Intact Cells Cy3NTAc Poly His  63 50-90 Plasma membrane protein, permeabilized cells  Cy5NTAc 43 34-...Intact Cells Cy3NTAc Poly His  65 52-94 Plasma membrane protein, permeabilized cells  Cy5NTAc 59 47-...
  5. Fluorescent Proteins 101: Luciferase

    Type
    Blog Post
    Published
    June 24, 2014, 3:59 p.m.
    ...such as excitation of Green Fluorescent Protein (GFP) because of the nature of the chemical reaction compared...to using a high-intensity laser to rapidly excite GFP. As a result of the different mechanisms to produce...as the most efficient means to disrupt the cell membrane and deliver the substrate; however, secreted luciferase...
  6. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...viral genomes (vg)/cell, pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] (Addgene 84445-AAVrg) alone at 1.1E6 vg/mL, ...prep # 55632-AAVrg). pAAV-CAG- FLEX-rc [Jaws-KGC-GFP-ER2] was a gift from Edward Boyden (Addgene viral...filtration through a 0.45 µm polyethersulfone (PES) membrane. The preparations are then aliquoted, frozen, ...
  7. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    Published
    March 3, 2020, 2:15 p.m.
    ...levels similar to that of a single AAV expressing GFP. The lab tested the base editors in vivo and achieved...voltage indicators, however, fall short compared to GFP-based sensors in terms of response and brightness...-opsin GEVIs. With an increase in the neuron’s membrane potential, Ace absorbance increases and results...
  8. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    Published
    April 24, 2015, 2:39 p.m.
    ...proteins, which are directly trafficked to cell membranes. Membrane trafficking of farnesylated derivatives enables...fluorescent proteins: RFP (red), YFP (yellow) and M-CFP (membrane tethered cyan). Without Cre recombination, RFP...-2.1 construct can express one of four colors (n-GFP, RFP, YFP or M-CFP.) The construct contains two tandem... and sequence overlap (coral mOrange2, jellyfish EGFP, and sea anemone mKate2.) They then successfully... is retained in Brainbow-3.0, but with mOrange2, EGFP and mKate2 as the fluorophores; mOrange2 is the ...
  9. Hot Plasmids February 2024

    Type
    Blog Post
    Published
    Feb. 20, 2024, 2:45 p.m.
    ..., Myc and SV40 Nuclear Localization Signals, and GFP. B) Workflow to assess Cas9-PAGE editing with viral...variants:   Figure 2: Inner mitochondrial membranes imaged with structured illumination microscopy...2023). StayGold variants for molecular fusion and membrane-targeting applications. Nature Methods, 1–9. https...
  10. The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins

    Type
    Blog Post
    Published
    July 29, 2014, 6:16 p.m.
    ...from Joachim Goedhart: Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection.  ...F-Actin to mitochondria, from the nucleus to cell membrane, this collection has fluorescently tagged versions...sun, from the standard oldies but goodies (e.g., EGFP and YFP) to the new and improved fruit colors (e.g...
  11. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 4, 2017, 2:30 p.m.
    ... of vesicle fusion and release from the plasma membrane by fusing various vesicle cargoes to FP timer ...fusing Medium-FT to LAMP-2A, a lysosome-associated membrane protein, and following its subcellular localization...Fluorescent Protein Aggregation Catch Up On Your GFP Basics Get Advice on Choosing Your Fluorescent Protein...
  12. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ...fluorescent protein that could be co-imaged with GFP using the same wavelength for excitation. The result...optogenetic accuators to depolarize or hyperpolarize membranes. Traditional FRET reporters could not be used ...Venus fluorescent protein in conjunction with a membrane-inserted voltage sensing domain (LOTUS V; Luminescent...Universal Sensing of Voltage). With this sensor, membrane depolarization increases the BRET signal whereas...
  13. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    Published
    May 15, 2018, 12:43 p.m.
    ...plasmids described in this work including the soCoChR-GFP plasmid as well as several derivatives encoding alternative... protein kinase A (PKA) activity at the plasma membrane in living cells using a construct in which TagRFP-T...ShadowG, a novel dark green fluorescent protein (GFP) with an absorption peak of 486 nm and ShadowY, a...
  14. Year of the Camelid: Antibody Style

    Type
    Blog Post
    Published
    May 9, 2024, 1:15 p.m.
    ...highlighted back in 2015, where sdAbs were used to create GFP scaffolds for transcriptional activation.  sdAbs ...Llamas Required - Synthetic Nanobodies Against Membrane Proteins More resources from addgene.org  Plasmids...
  15. Hot Plasmids - October 2022

    Type
    Blog Post
    Published
    Oct. 6, 2022, 1:15 p.m.
    ...photoactivation of HcKCR1 in mouse neurons. D) Membrane voltage traces of action potentials in a HcKCR1...Addgene’s western blot for the myc-tagged protein GFP-smFP-myc expressed from Plasmid 98926. See the Applications...
  16. Donations from Addgene to Yield Answers for Rare Disease Researchers

    Type
    Blog Post
    Published
    Sept. 15, 2015, 2:30 p.m.
    ...play a role in many cell processes involving cell membrane dynamics. When they learned there was a disease... University with David Goldstein’s lab, will use GFP-ATP1A3, FLAG-ATP1A3, and ATP1B3 plasmids to explore...
  17. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    Published
    May 18, 2021, 1:15 p.m.
    ... hSyn FLEx mGFP-2A-Synaptophysin-mRuby from Liqun Luo’s lab expresses membrane tethered GFP to enhance...  After validating this method through monitoring GFP degradation, this protein compartment was used to...targeted synaptophysin tagged eGFP. AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE from Hongkui Zeng's lab...
  18. Another Pathway into Cells: iTOP

    Type
    Blog Post
    Published
    June 23, 2015, 8:37 p.m.
    ...Tat. Although CPP fusions can translocate a cell membrane, the presence of the CPP may alter protein function...Neuronal stem cells transduced with iTOP resulting in GFP expression. Image courtesy of Diego D'Astolfo. ...
  19. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    Published
    Sept. 8, 2016, 2:30 p.m.
    ...chemistry! The discovery of green fluorescent protein (GFP) sparked a renaissance in biological imaging. Suddenly...advances lead to an obvious question: In this age of GFP and its ilk, why should cell biologists talk to chemists...analogs without sacrificing their small size and membrane permeability (Figure 2c). These cell-permeable...
  20. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    Published
    Dec. 6, 2022, 2:15 p.m.
    ...are generated by inserting a circularly-permuted GFP into an intracellular loop of various G-protein-coupled...they observe fluorescence localizes to the plasma membrane only when the RTK is activated and the ITAM phosphorylated...relies on a change in localization to the cell membrane and not a direct change in the fluorescent properties...devised a “Modular fluorescence unit fused with TRansmembrane region-to-IntrAcellular loop linkers” (MTRIA...
Showing: 21 - 40 of 51 results