Skip to main content
Addgene
Showing: 1 - 50 of 68 results
  1. Lentivirus Production

    Type
    Protocol
    ...drifted. Filter the solution through a 0.22 μm membrane. Aliquot 500–1000 μL into sterile tubes. Store...determine what ratio gives the highest percentage of GFP positive cells. Refer to the table below for a possible...Data Figure 1: 293T cells were transfected with the GFP-expression plasmid pRosetta using μg total DNA to...efficiencies as measured by the highest proportion of GFP positive cells without limiting cell growth. Left...Left panels: bright field images; right panels: GFP channel images. Last reviewed on: August 2, 2023...
  2. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    ...contacts that bring apposing membranes together and can wrap the membranes into stacked structures including... Partitioning of lipid-modified monomeric GFPs into membrane microdomains of live cells. Science, 2002...remain unaware that EGFP/GFP is prone to forming noncovalent dimers. This property of EGFP can lead to significant...contributed by guest blogger Erik L. Snapp. Stop using EGFP/GFP for fusion proteins! Despite multiple studies ...protein or Enhanced Green Fluorescent Protein (GFP/EGFP) for a transcriptional reporter or as a general...in the world, is not inert. In all seriousness, EGFP/GFP has a real nontrivial propensity to noncovalently...best solution is to use truly monomeric FPs. EGFP and other GFP family members (superfolder, Emerald, CFP...
  3. Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification

    Type
    Blog Post
    ...detect its co-localization with a membrane-localized mCherry or GFP protein. With this system, if the ...Lab monitored how AsLOV2 insertion into mCherry or GFP nanobodies affected binding of their target proteins...bind, it co-localizes with its target at the cell membrane and when the OptoNB can’t bind, it localizes to...more OptoNBs, one against mCherry and one against GFP. Insertion of AsLOV2 into loop 1 always generated....  AsLOV2 Inertion Site mCherry #1 mCherry #2 GFP Loop 1 dark-induced dark-induced dark-induced ...proliferation and differentiation.  Since Ras is a membrane localized GTPase, the team fused a mCherry OptoNB...OptoNB-SOS fusion protein in cells that expressed membrane-localized mCherry. With darkness, SOS binds Ras...
  4. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    ...protein of interest. The green fluorescent protein (GFP) and mCherry, two of the most common tags currently...only a small proportion were found on the plasma membrane (top panel in Fig. 1). Of course, this was problematic...DREADD receptors need to be located on the plasma membrane to be activated by the DREADD actuator after it.... Although we found a larger proportion on the membrane in mice neurons, most of them were still cytoplasmic...limited expression of hM4Di‐mCherry at the plasma membrane in monkey neurons, we next hypothesized that a...improve the transport of this DREADD to the plasma membrane. The haemagglutinin (HA) tag is a small 9 amino...resulted in abundant expression of hM4Di in plasma membranes in both species (bottom panel in Fig. 1). Our ...
  5. Visualizing Translation at the Single Molecule Level

    Type
    Blog Post
    ...switching, TRICK requires the following components: a GFP protein that binds to the coding region of a reporter...reporter mRNA via a PP7 coat protein (NLS-PCP-GFP) an RFP protein that binds to the 3’ UTR of the reporter...Plasmid 64542 or 64543) Before translation, both GFP and RFP are bound to the reporter, causing it to ...appear yellow. The act of translation pushes NLS-PCP-GFP off of the mRNA, leaving only a red FP bound to the... contains a nuclear localization signal, NLS-PCP-GFP then returns to the nucleus to bind cognate untranslated...verified that translation is necessary for removal of GFP from the TRICK reporter. Delayed translation is known...Oskar protein levels were negatively correlated with GFP intensity, showing that TRICK is an accurate readout...
  6. New and Upcoming Viral Vectors - Spring 2019

    Type
    Blog Post
    ...-KGC-GFP-ER2) and cre-dependent version (pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2]) of this membrane-embedding...pAAV.hSyn-FLEX.iGABASnFR.F102G pAAV-Syn-Archon1-KGC-GFP-ER2 pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2] pAAV-FLEX-tdTomato pAAV-GFAP104...pAAV-mDlx-NLS-mRuby2 CAG-NLS-GFP pAAV-hSyn-mCherry Read on to learn more!  Calcium sensor AAV The GFP-based calcium...37825  AAV8*, AAV9*  pAAV-CAG-GFP 50465  AAV8*, AAV9*  pAAV-hSyn-EGFP *Also coming in our new 20 ...GABA sensor (pAAV.hSyn-FLEX.iGABASnFR) has good membrane localization and brightness. The F102G variant...pAAV-GFAP104-mCherry, pAAV-mDlx-NLS-mRuby2, CAG-NLS-GFP,and pAAV-hSyn-mCherry at Addgene. Viral vectors coming...IRES EGFP Chimeric channels for neuronal manipulation 119742  AAV5  AAV SYN PSAM4 GlyR IRES EGFP Chimeric...
  7. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ...specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID along with...cell membrane isn’t powerful enough, what about altering the chemical composition of the membrane itself...SuperPLD hydrolyzes membrane PC to PA. A) OptoPLD is recruited to plasma membrane or lysosomes and its...phosphatidic acid probe GFP-PASS. B) Time series of HEK 293T cells expressing plasma membrane-targeted SuperPLDx30...optogenetic tools to control the cytoskeleton and cell membrane dynamics. First, a quick refresher.  The tools...undergoes a conformational change and binds the plasma membrane under blue light CRY2-CIB1 heterodimerizes or .... The Chow Lab has also generated a variety of membrane-recruitment optogenetic tools for manipulating...
  8. Sensing Neuronal Dopamine

    Type
    Blog Post
    ...sensor was engineered from a GFP variant and a DA receptor such that GFP fluoresces when the receptor ...on a similar strategy: a GFP variant bound to a DA receptor, such that the GFP fluoresces upon DA-receptor...performed with a probe surrounded by a semipermeable membrane, which is inserted into the region of the brain...passive diffusion of DA across the semipermeable membrane from the extracellular fluid into the perfused...
  9. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    ...protein) associates with autophagosomal membranes, so tagging it with GFP allows for tracking the localization...Autophagosomes are relatively easy to identify: double-membraned structures containing undigested cytoplasmic contents...electron microscopy analysis is time consuming. GFP-tagged Atg8p or LC3: ATG8 is part of a group of genes...autophagosomes and autophagic bodies in yeast. A GFP-tagged version of LC3, the mammalian Atg8p homolog...pH-insensitive RFP variant, DsRed.T3 and a pH-sensitive GFP variant, super ecliptic pHluorin (SEP). They are ...Fluorescent Protein Oligomerization Seeing Red: Simple GFP Photoconversion Light Sheet Fluorescence Microscopy...
  10. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    ...three different membrane-targeting flavors – PDGFR, GPI, and SGZ, to improve neuronal membrane trafficking...regulation. To create iGluSnFR, the Looger lab wedged GFP between two halves of a bacterially derived periplasmic...increase in fluorescence intensity from the inserted GFP. After characterizing iGluSnFR in vitro, showing ...segment (PDGFR TMD) that anchors the sensor to the membrane and displays it on the extracellular side. In ...generation of iGluSnFR, the Looger lab swapped eGFP for sfGFP to create SF-iGluSnFR. This improved variant.... PDGFR is a C-terminal fusion to the PDGFR transmembrane domain in the mammalian expression pMinDisplay...glycosylphostidylinositol anchor. SGZ contains a PDGFR transmembrane domain, followed by a modified form of the cytosolic...
  11. Fluorescent Biosensors for Measuring Autophagic Flux

    Type
    Blog Post
    ...Addgene! mRFP-GFP-LC3 tandem fluorescent protein This autophagy biosensor contains mRFP and GFP fused to the...flux. mRFP-GFP-LC3 functions as an autophagy bionsensor via the same mechanism as Rosella: GFP is quenched...decrease in red puncta. Find the mRFP-GFP-LC3 plasmid at Addgene! GFP-LC3-RFP-LC3∆G: A fluorescent probe ...autophagy biosensor is the GFP-LC3-RFP-LC3ΔG probe. It’s similar to the mRFP-GFP-LC3 Tandem Fluorescent Protein...molecule of GFP-tagged LC3 for every one molecule of RFP-tagged LC3∆G. Like the unmodified LC3, GFP-LC3 is ...Autophagic flux is estimated by the GFP/RFP ratio: a decrease in GFP fluorescence relative to RFP fluorescence...variant, DsRed.T3, that’s connected to a pH-sensitive GFP variant, super ecliptic pHluorin (SEP). In the absence...
  12. SunTag and Fluorescent Imaging

    Type
    Blog Post
    ...al. observed some GFP aggregation, which they reduced by using superfolder GFP (sfGFP) with the small solubility...scaffold fully occupied with GFP has a molecular weight of 1,400 kDa vs 24 kDa for GFP alone. Although traditional...nomenclature throughout the rest of this blog post, GFP refers to sfGFP-GB1.   Tanenbaum et al. examined the power...finding that plasma membrane-targeted CAAX-SunTag was 18-fold brighter than sfGFP! The high SunTag signal... copies of the short epitope GCN4. GCN4 recruits GFP fused to the cognate scFV antibody, which is expressed...physiology. Having seen that mitochondrial tracker GFP-mitoNEET can impair mitochondrial function, they ...they examined the effects of mitoNEET-SunTag-GFP. As expected, they obtained bright images of mitochondria...
  13. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    ... from the default GFP (green) to RFP, CFP, or YFP fluorescence. In Brainbow 2.1, GFP is tagged with a ... cells that randomly express either nuclear GFP, membrane CFP, cytoplasmic YFP, or cytoplasmic RFP in ... the fluorophores RFP (red), YFP (yellow), and membrane-tethered CFP (cyan). Brainbow 1.1 adds OFP (orange...and activate fluorescence of the downstream OFP, GFP, or RFP. Cells that express the construct but do ...control of Flp recombinase. The XFPs in Flybow are membrane-tethered, making this system useful for visualization...incorporates the highly photostable fluorophores mOrange2, EGFP, and mKate2 (red) to solve the problematically low...
  14. Fluorescent Protein Guide: Subcellular Localization

    Type
    Collection
    ... Gerlich 25999 LV-GFP Chromatin H2B GFP Elaine Fuchs 11680 H2B-GFP Chromatin H2B GFP Geoff Wahl 21210 ...Mitochondria MCU GFP Vamsi Mootha 49153 GFP-Mff Mitochondria-Outer Membrane Mff acGFP1 Gia Voeltz 158007...61803 GFP-Rab7A Late endosomes Rab7a AcGFP Gia Voeltz 12605 GFP-rab7 WT Late endosomes RAB7 GFP Richard...37351 pQC membrane TdTomato IX Membrane Palmitoylation TdTomato Connie Cepko 22479 FUmGW Membrane Palmitoylation...Mitochondria-Outer Membrane TOMM20 mCherry* Michael Davidson 118736 pBOB-CARMIL2 BH domain-GFP Plasma Membrane CARMIL2...Palmitoylation GFP Connie Cepko 14757 pCAG-mGFP Membrane Palmitoylation sequence from GAP43 EGFP Connie Cepko...Cepko 61099 Lck-GFP Membrane Palmitoylation sequence from Lck EGFP Steven Green 36209 pPalmitoyl-mTurquoise2...
  15. General Transfection

    Type
    Protocol
    ...drifted. Filter the solution through a 0.22 µm membrane. Aliquot 500–1000 µL into sterile tubes. Store...determine what ratio gives the highest percentage of GFP positive cells. Refer to the table below for a possible...
  16. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ...to endogenously tag a wide variety of genes with GFP. These plasmids are available through Addgene, and...is the addition of a useful coding sequence (i.e. GFP) into a gene while leaving other features of the ...we show an example gene targeted for tagging with GFP at the C terminus, resulting in one edited allele...allele, or gene copy. The coding sequence for GFP is transcribed and translated along with the gene, producing...shown in the example above, we inserted a linker and GFP tag at the end of the last exon of the gene to ensure...matching the gene of interest, a linker, and the GFP tag sequence to be inserted. Created with BioRender.com...Cas9 makes at the target site. The whole segment—a GFP tag and 1kb of homologous DNA flanking both sides...
  17. Fluorescent Proteins 101: Fluorescent Biosensors

    Type
    Blog Post
    ...biosensors as an example. Many calcium biosensors include GFP fused to calmodulin (CaM) and the chicken myosin ...characteristics like pH, redox state, and voltage across membranes. These sensors generally follow the same pattern...endoplasmic reticulum (ER), vacuole or cellular membranes. Examples include blue flame mitochondria-targeted...an organelle-specific targeting signal or a transmembrane domain, biosensors can be targeted to a particular...
  18. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    ...hiPSCs  EMX1  67% Mouse BM cells from GFP transgenic mouse  GFP  ~60-65% Human hepatocytes  Myd88  ~...spontaneously assemble into particles at the cell membrane. Mature Nanoblades then bud into the cell culture... where they can then fuse with the target cell membrane, and transfer their CRISPR cargo into the zygote...
  19. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...GTPase in division) GFP-Mff (outer membrane protein) Late Endosome mCherry-Rab7A GFP-Rab7A     ...both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins...and plasma membrane reporters as well as secretory proteins or proteins targeted to membrane sub-domains...Compartment mCherry-Rab5 BFP-Rab5 GFP-Rab5B Vacuolar & Budding Compartment GFP-Rab4B Mitochondria mito-BFP...protein Knock-off, is made up of three components: a GFP protein that binds to the coding region of a reporter...reporter mRNA via a PP7 coat protein (NLS-PCP-GFP) a RFP protein that binds to the 3’ UTR of the reporter...these dCas9s with one of three fluorescent proteins (GFP, BFP and mCherry), the authors were able to visualize...
  20. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    ...Favorite Fluorescent Protein Oligomerizes in Cells GFP oligomerization can lead to misleading artefacts ... your research, check out this post! R Bodies: Membrane-Rupturing Microscopic Tools In this post, Jessica...also showed that R-bodies can rupture E. coli membranes. Read the full post for potential applications...
  21. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

    Type
    Blog Post
    ... AAV2 interacts with the packaging cell's cell membrane When packaging adeno-associated viral vectors ...supernatant, the viral particles associate with the cell membrane and are taken into the cell via endocytosis. In...endocytosis of a fluorescent dye and transduction of GFP-expressing AAV particles we did not observe any difference...
  22. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...S65T) in the original jellyfish GFP (AvGFP) to create enhanced GFP (EGFP), a variant with improved brightness... There are numerous GFP variants. Here I use the name ‘GFP’, which refers to mEGFP. This variant was made...examined on a case-by-case basis. GFP is sensitive to acid The chromophore of GFP can exist in a protonated and...its acid tolerance is maintained in cells.   GFP is big GFP is a 28 kDa protein that resembles a cylinder...2013; Hilaire et al., 2017). GFP can only be used for tagging proteins GFP fusion proteins are made by ... a fusion with GFP allows for detection of genomic locations (Chen et al., 2013). GFP fluorescence overlaps... al., 2018). GFP is not tolerated in my fusion It is standard practice to attach the GFP to the N- or ...
  23. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...dim GFPs The availability of the three dimensional structure of GFP, its brighter mutant S65T-GFP, and...and GFP shRNA. Mice carrying the R26DsRedR; CRUSH-GFP; and Nestin-Cre alleles showed efficient GFP knockdown...identified a family of GFP proteins in the cephalochordate Branchiostoma floridae, named bfloGFPs. They went on...and demonstrate that they can successfully create GFP fusion proteins with a variety of genes across the...enabled scientists to engineer a wide variety of GFPs with diversity in fluorescence brightness, intensity...relies on several observations: Rapamycin is a membrane-permeable pharmacological agent that can rapidly...extraneous proteins bound to its outer mitochondrial membrane. Sequestration is accomplished by knocking down...
  24. Hot Plasmids - August 2020

    Type
    Blog Post
    ...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...
  25. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    ...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...
  26. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...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...
  27. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...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-...
  28. Fluorescent Proteins 101: Luciferase

    Type
    Blog Post
    ...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...
  29. Viral Production at Addgene

    Type
    Blog Post
    ... 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, ...
  30. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...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...
  31. Fluorescent Protein Guide: Biosensors

    Type
    Collection
    ...Calcium erGAP3 (GFP-Aequorin Protein) for imaging of Ca+ dynamics in endoplasmic reticulum GFP-Aequorin Protein...FlincG3 (GFP-based cGMP sensor) for imaging in C. elegans neurons Using a Robust and Sensitive GFP-Based ...trafficking to the plasma membrane Acute disruption of the synaptic vesicle membrane protein synaptotagmin...measurements of membrane charges A FRET sensor enables quantitative measurements of membrane charges in live...2020 GENIE Project Calcium jGCaMP7 High-performance GFP-based calcium indicators (Constitutive or Cre-dependent...sensor iATPSnFR1.0 and iATPSnFR1.1 (cytoplasmic, membrane-displayed, expression in neurons, astrocytes) ...cyclic AMP) Signaling reporter island (SiRI) with GFP-based fluorescent reporter cAMPr Spatial Multiplexing...
  32. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...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 ...
  33. Hot Plasmids February 2024

    Type
    Blog Post
    ..., 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...
  34. The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins

    Type
    Blog Post
    ...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...
  35. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ... 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...
  36. AAV Molecular Tools

    Type
    Collection
    ...Cre-dependent Cre-dependent expression of membrane-bound GFP and synaptophysin-mRuby for labeling of axon...System Activity Serotype PI 124364 pAAV-FLEX-DTR-GFP CBA-driven, Cre-dependent Cre-dependent expression...expression of diphtheria toxin receptor fused to GFP for studying cell ablation. 2 Jessell , Azim 45580 pAAV-... Serotype PI 98747 pAAV-FLEX-EGFPL10a EF1a-driven and Cre-dependent EGFP-tagged ribosomal L10a 5, rg* ...and synaptophysin-EGFP for labeling of axon terminals. 1 Zeng 71760 pAAV hSyn FLEx mGFP-2A-Synaptophysin-mRuby...Serotype PI 51509 AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE Syn-driven, Cre-dependent Cre-dependent expression...
  37. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...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...
  38. Year of the Camelid: Antibody Style

    Type
    Blog Post
    ...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...
  39. Hot Plasmids - October 2022

    Type
    Blog Post
    ...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...
  40. Antibody Plasmid Collection

    Type
    Collection
    ...making synthetic nanobodies (sybodies) against membrane protein targets. Synthetic single domain antibodies...antibodies for the conformational trapping of membrane proteins. Elife. 2018 May 24;7. and Generation of...find antibody plasmids for: Common antigens such as GFP or mCherry Monoclonals, Nanobodies, Sybodies, or ...peptide, from the popular SunTag system, fused to sfGFP for imaging. Learn more about antibodies and their...
  41. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ... 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...
  42. Another Pathway into Cells: iTOP

    Type
    Blog Post
    ...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. ...
  43. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    ...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...
  44. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ...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...
  45. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...nucleocapsid protein (Plasmid #141391-LV [+N]) or membrane protein (Plasmid #141386-LV [+M], negative control...Figure 5: A) Representative confocal images showing GFP expression from ssAAV5s with hIBA1 or hIBA1a promoters...Quantifications showing high microglia specificity of GFP expression for the indicated ssAAV5s. C) Quantifications...
  46. New England Biolabs Cell-Imaging Plasmid Collection

    Type
    Collection
    ...comprehensive comparison to GFP, please refer to NEB's comparison of SNAP-tag, CLIP-tag, and GFP . Technology SNAP-tag...with a recognizable sub-cellular localization (membrane, nucleus, etc.) are also available Non-toxic to...pSNAP-CaaX Control Plasmid SNAP-tag Control for endomembrane (CaaX box) localization 101129 pSNAPf-Cox8A ...
  47. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    ...line, Hep3B. From left to right: Mitotracker Red, GFP for the biosensor, Hoechst, and overlay).   ...multiplexed imaging of neurotransmitter, calcium, and membrane voltage,” says Tian. Figure 5: Citrine-tagged...(DKFZ) who has developed new redox sensitive GFP2 (roGFP2)-based biosensors for live cells. His work focuses..., Pedre Pérez’s biosensor fuses MPST to roGFP2. Because roGFP2 contains two cysteines near the chromophore...the MPST hydrosulfide can be transferred to the roGFP2 cysteine thiols and change its fluorescence properties...
  48. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    ...sensors available that will localize at the plasma membrane. GCaMP-X is a nucleus-targeted GCaMP6 with an ...ones? Many of the available biosensors use a single GFP or CFP/YFP-derived FRET pair. If you need to free...
Showing: 1 - 50 of 68 results