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  1. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    ...signal. Prior to the development of these probes, fluorophore binding aptamers and MS2-binding fluorescent ...aptamers or MS2 tags using the same number of fluorophores. You can find plasmids containing the cobalamin...expresses eGFP to indicate reporter abundance, and mCherry fluorescence is activated when APOBEC-Cas9n-UGI-mediated...activity edits out a frame-shift mutation within the mCherry reporter. Quantification of editing efficiencies...efficiencies simply involves tracking the ratio of mCherry-positive to eGFP positive cells through fluorescence...
  2. New and Upcoming Viral Vectors - September 2019

    Type
    Blog Post
    ...pAAV-hSyn-hChR2(H134R)-mCherry (26976-AAV5) pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP (112677-AAV5) ...scientists access to a wide array of promoters and fluorophores for validating and testing purposes. We will...GABAergic interneurons. pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP (112677-AAV5 and 112677-AAVrg): The EF1a ... a Cre-dependent switch that expresses nuclear mCherry in the absence of Cre, and expresses nuclear EGFP...DreO-bGHpA (50363-AAV5) Flp vectors pAAV-EF1a-mCherry-IRES-Flpo (55634-AAV1) pAAV-EF1a-Flpo (55637-AAV1...
  3. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    ...expression of transgenes of interest. Furthermore, fluorophore variants (for example green and red fluorescent...interest. The green fluorescent protein (GFP) and mCherry, two of the most common tags currently used, are...used a construct in which the hM4Di was fused to mCherry. To investigate the localization of the hM4Di, ..., we used primary antibodies against mCherry, and secondary antibodies coupled to nanogold particles, ...proteins Given the limited expression of hM4Di‐mCherry at the plasma membrane in monkey neurons, we next...small change made a big difference. Replacing the mCherry with the HA tag resulted in abundant expression... of the transgene of interest. In our studies, mCherry interfered with the transport to the membrane of...
  4. Recombinases AAV Preps

    Type
    Collection
    ...none rg* Wickersham 55634 pAAV-EF1a-mCherry-IRES-Flpo EF1a mCherry (not a fusion tag) 1, rg* Deisseroth..., 2, 5, 8, 9 Wilson 55632 pAAV-Ef1a-mCherry-IRES-Cre EF1a mCherry (not a fusion tag) 8, rg* Deisseroth... Syn none 9 Wilson 107312 AAV-hSyn-mCherry-P2A-Cre-WPRE Syn mCherry 1 Yang 107738 pAAV-hSyn-Cre-P2A-dTomato...control gene expression. Dre AAV ID Name Promoter Fluorophore Serotype(s) PI 50363 AAV phSyn1(S)-DreO-bGHpA...Syn none 5, rg* Zeng Flpo AAV ID Name Promoter Fluorophore Serotype(s) PI 51669 AAV phSyn1(S)-FlpO-bGHpA...none 1, rg* Janelia VCre AAV ID Name Promoter Fluorophore/Tag Serotype(s) PI 55638 pAAV-EF1a-vCre EF1a ...none 8, rg* Deisseroth Cre AAV ID Name Promoter Fluorophore/Tag Serotype(s) PI 105551 pENN.AAV.CamKII.HI....
  5. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ...specifically drives expression of mCherry instead of BFP. Beyond switching fluorophores, why is FLEx useful? The...that turns BFP expression off, while turning on mCherry expression. For this FLEx switch to successfully...sequence in the sense orientation, followed by the mCherry coding sequence in the antisense orientation (Figure... switch will express BFP in the absence of, or mCherry in the presence of Cre recombinase.   Once ...
  6. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ... you'll need to pick a fluorophore for each target and ensure the fluorophores, together, are able to ...outlining a few commonly used fluorophores and their emission spectra. Some fluorophores carry their emission ...and RB545. Fluorophores can also come in combination of two conjugated single fluorophores, like APC-Cy7...respective colors of a few commonly used fluorophores. Each fluorophore has a defined emission maximum. Note...individual fluorophores. Table 1 describes the brightness of a few commonly used fluorophores. A fluorophore's...voltage, the concentration of the fluorophore, and whether that fluorophore appears alone or in conjugation...conjugation with another fluorophore. The brighter a fluorophore, the better the signal-to-noise ratio, and the...
  7. HA Frankenbody, a New Imaging Tool to Visualize Single Molecules and Nascent Peptides

    Type
    Blog Post
    ...scFvs) (Inbar et al., 1972) are fused to mature fluorophores, and can bind and light up epitope tagged proteins...colocalization of GFP tagged HA frankenbody with an mCherry, 4xHA tagged histone protein (H2B) in U2OS cells... other fluorescent proteins and tags including mCherry, HaloTag and SNAP-tag for multi-color imaging. ...
  8. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ... are named for soup ingredients! From pSOUP to mCherry, satisfying ingredients reagents permeate the molecular...known promoter. Addgene’s Plasmids 101: Aptamer Fluorophores describes the enrichment process used to evaluate...hydroxybenzylidene-imidazolinone-2-oxime (DFHO), a fluorophore modified from DsRed. DFHO shares many structural...In addition to these established RNA aptamer fluorophores, more are in development, including Carrot and...Dolgosheina, Elena V., et al. "RNA mango aptamer-fluorophore: a bright, high-affinity complex for RNA labeling...polymerase III transcription using a photostable RNA–fluorophore complex." Nature chemical biology 13.11 (2017...on the Addgene blog Learn more about aptamer fluorophores Investigate other fluorescent biosensors Find...
  9. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...), tagRFP (pKa = 3.1) and mCherry (pKa = 3.8). We have observed that mCherry (and mTurquoise2, not shown... bind specifically and with high affinity to fluorophores. Examples are the FlAsH tag, the His tag and...enzyme to covalently label the peptide with a fluorophore. Both systems are a hybrid between a genetically...
  10. Control AAV Preps

    Type
    Collection
    ...AAV-EF1a-BbChT EF1a mCherry and mTFP Cre dependent 9 Sanes 114471 pAAV-Ef1a-fDIO mCherry EF1a mCherry Flp dependent...Constitutive 5 Zeng 58909 pAAV-GFAP104-mCherry GFAP104 mCherry Constitutive 5 Boyden 59462 pAAV-CAG-tdTomato...Constitutive 2 Patrick 114469 pAAV-CaMKIIa-mCherry CaMKIIa mCherry Constitutive 1, 2, 5, 8, 9, rg*, PHP.eB...PHP.eB Deisseroth 114470 pAAV-Ef1a-mCherry EF1a mCherry Constitutive 1, 2, 5, 8, 9, rg*, PHP.eB Deisseroth...Deisseroth 114472 pAAV-hSyn-mCherry hSyn mCherry Constitutive 1, 2, 5, 8, 9, rg* Deisseroth 117382 hSyn1-eYFP ...112677 pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP EF1a NLS-mCherry or nls-EGFP Cre dependent 1, 2, 5, ...1, 2, 5, 8, rg* Roth 50459 pAAV-hSyn-DIO-mCherry hSyn mCherry Cre dependent 1, 2, 5, 8, 9, rg* Roth 51502...
  11. MXS Chaining

    Type
    Blog Post
    ... partner, which directed localization of the fluorophore, and thus enabled labeling of particular structures...with strong expression and detection of each fluorophore at the respective subcellular structure (Figure...for Labeling Cellular Structures Construct Fluorophore Excitation/emission maxima Tethering partner ...the tyrosine-protein kinase Lyn) Membranes 3 mCherry 587nm/ 610 nm human β-Actin Actin 4 Citrine 516nm...
  12. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ...the intestine. Figure 3: Expression of fluorophore-tagged GOLG-4 through eight stages of embryonic...traditional homologous recombination methods). By using mCherry for selection of sgRNA plasmid transformants, they...with the sgRNA cassette on a plasmid expressing mCherry and resistance selection marker HygR. The CRISPR...disruption of the target gene. Lastly, curing the mCherry-sgRNA plasmid leaves an edited strain carrying ...
  13. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ... ER mCherry-Sec61 beta BFP-Sec61 beta BFP-KDEL Rtn4a-GFP (tubular ER) Microtubules mCherry-alpha-tubulin...mito-BFP mCherry-Drp1 (GTPase in division) GFP-Mff (outer membrane protein) Late Endosome mCherry-Rab7A... fluorescent pH sensors, a set of luciferase-fluorophore pH fusion reporters, pRSETb-pHlash and pcDNA3.1...photon energy is transferred to the acceptor fluorophore, a circularly permuted Venus. In the case of ...of pHlash, the signal from the fluorophore is pH-dependent. Signal is simultaneously acquired from the ...Pre-constructed Entry vectors containing Cas9, EGFP, mCherry, iRFP, tdTomato, luciferase, LacZ, puromycin or...one of three fluorescent proteins (GFP, BFP and mCherry), the authors were able to visualize simultaneously...
  14. Deisseroth INTRSECT Collection

    Type
    Collection
    ...55632 pAAV-Ef1a-mCherry-IRES-Cre None 55637 pAAV-EF1a-Flpo None 55634 pAAV-EF1a-mCherry-IRES-Flpo None ...137132 pAAV-Ef1a-Con/Fon-mCherry Cre AND Flp 137133 pAAV-Ef1a-Con/Foff 2.0-mCherry Cre AND NOT Flp FRT/F5...pAAV-nEF-Con/Fon-ChR2-mCherry Cre AND Flp 137143 pAAV-nEF-Con/Foff 2.0-ChR2-mCherry Cre AND NOT Flp FRT/...pAAV-nEF-Coff/Fon DREADD Gi-mCherry Flp AND NOT Cre 177672 pAAV-nEF-Con/Fon DREADD Gi-mCherry Cre AND Flp 177673...pAAV-nEF-Con/Foff DREADD Gi-mCherry Cre AND NOT Flp 183532 pAAV-nEF Con/Fon DREADD Gq-mCherry Cre AND Flp 183533...Con/Foff DREADD Gq-mCherry Cre AND NOT Flp 183534 pAAV-nEF Coff/Fon DREADD Gq-mCherry Flp AND NOT Cre Dual...configuration have been developed, including additional fluorophores, excitatory and inhibitory opsins , genetically-encoded...
  15. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    ...mutagenesis screening for enhanced RFP variants. PA-mCherry (E26V / A58T / K69N / L84F / N99K / S148L / I165V...UV-Violet or blue Green/ Red 4,500 High 553/ 573 PA-mCherries Monomer UV-Violet Dark/ Red >3,000 Medium 570/... of the technique relies heavily on the same fluorophore being imaged multiple times in order to reconstruct.... 2.Subach, Fedor V., et al. "Photoactivatable mCherry for high-resolution two-color fluorescence microscopy...
  16. Optogenetics AAV Preps

    Type
    Collection
    ...-ChR2-mCherry nEF ChR2 mCherry Cre dependent 8 Deisseroth 137144 pAAV-nEF-Coff/Fon-ChR2-mCherry nEF ChR2...1, 9 Svoboda 83898 pAAV-mDlx-ChR2-mCherry-Fishell-3 Dlx ChR2 mCherry Constitutive 1, 9, rg* Fishell 135634...135634 pAAV-S5E2-ChR2-mCherry nEF ChR2 mCherry Constitutive 1, 9, PHPeB Dimidschstein 137142 pAAV-nEF-...pAAV-nEF-Con/Fon-ChR2-mCherry nEF ChR2 mCherry Cre and Flp dependent 8 Deisseroth 137143 pAAV-nEF-Con/Foff 2.0...pAAV-EF1a-double floxed-hChR2(H134R)-mCherry-WPRE-HGHpA EF1a ChR2/H134R mCherry Cre dependent 1, 5, 8, 9, rg*...Deisseroth 26975 pAAV-CaMKIIa-hChR2(H134R)-mCherry CaMKII ChR2/H134R mCherry Constitutive 1, 5, 9, rg* Deisseroth...Deisseroth 26976 pAAV-hSyn-hChR2(H134R)-mCherry Syn ChR2/H134R mCherry Constitutive 5, 8, 9 Deisseroth 28017...
  17. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ... al., 2007) and includes four promoters, six fluorophores with nonoverlapping emission spectra (N- and...reporter activation and silencing of factor-linked mCherry). They additionally discovered that monocistronic...last month, Robert Campbell’s lab added two new mCherry variants to the repository. These variants can ...ONE paper for more on the directed evolution of mCherry and the spectral properties of various RFPs. For...Listen to Our Podcast Segment on PhoCl LSSmCherry1 & RDSmCherry1: Engineering and directed evolution of...influence of structure on an FP’s properties. pBAD-LSSmCherry1 is a long Stokes shift variant, which could be...two-photon microscopy using Ti-Sapphire lasers. pBAD-RDSmCherry1 is a red-shifted variant which, with further ...
  18. Brain Initiative Collection

    Type
    Collection
    ...Gordon Fishell 83898-AAV1 pAAV-mDlx-ChR2-mCherry-Fishell-3 ChR2-mCherry fusion expression in forebrain GABA-ergic...Gordon Fishell 83898-AAV9 pAAV-mDlx-ChR2-mCherry-Fishell-3 ChR2-mCherry fusion expression in forebrain GABA-ergic... Fishell 83898-AAVrg pAAV-mDlx-ChR2-mCherry-Fishell-3 ChR2-mCherry fusion expression in forebrain GABA-ergic...135634-AAV1 pAAV-S5E2-ChR2-mCherry AAV vector to drive the expression of ChR2-mCherry in PV cortical interneuronsunder...135634-AAV9 pAAV-S5E2-ChR2-mCherry AAV vector to drive the expression of ChR2-mCherry in PV cortical interneuronsunder...-PHPeB pAAV-S5E2-ChR2-mCherry AAV vector to drive the expression of ChR2-mCherry in PV cortical interneuronsunder...expression. Bryan Roth 75033-AAV1 pAAV CD68-hM4D(Gi)-mCherry Expression of hM4D(Gi) using the CD68 promoter....
  19. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...p2A-ferritin-p2A-mCherry Mammalian Expression 74308 pcDNA3.0-Magneto2.0-p2A-mCherry Mammalian Expression...expression constructs fused with epitopes or fluorophores, generate expression vectors with various promoters...and named it Magneto 2.0. Using Magneto2.0-p2A-mCherry constructs, the lab verified that Magneto2.0 was...Expression 74334 pCR8-Magneto2.0-p2A-mCherry Gateway 74333 pCR8-Magneto2.0 Gateway 74307 pAAV-CMV-DIO-Magneto2.0...AAV 74302 pDestTol2CG2-Neurog1-Magneto2.0-p2A-mCherry-pA Zebrafish Expression   Human kinase domain...addition, some versions of the mAID vectors come with mCherry2 or mClover fluorescent proteins allowing you to...
  20. Fluorescent Protein Guide: Biosensors

    Type
    Collection
    ...sensors and separate nuclear localized dTomato fluorophore Ting Lab GCaMP6 Plasmids 2014 Jonathan Ting Calcium...biosensor for monitoring cAMP dynamics Single-fluorophore biosensors for sensitive and multiplexed detection...expression of fluorescent NADH/NAD+ biosensor Peredox-mCherry in plants In Vivo NADH/NAD+ Biosensing Reveals .... Luis Felipe Barros Pyruvate Pyronic single-fluorophore sensor for pyruvate A highly responsive pyruvate...
Showing: 1 - 20 of 20 results