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Showing: 1 - 20 of 59 results
  1. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    Published
    July 7, 2020, 1:15 p.m.
    ...three inactivated eGFP reporters- eGFP L202, eGFP L138, and eGFP L93. The modified eGFPs were placed downstream...comparing cells expressing only eGFP expression to cells expressing eGFP and mCherry. ACE was used to identify... To create the eGFP reporters the Harris lab individually mutated three codons in eGFP to eliminate fluorescence...editisome complexes. The eGFP reporters rely on the correction of point mutations in eGFP that results in the...mutated mCherry and a downstream constitutively active eGFP. To create the inactive mCherry, the lab introduced...fluorescence to monitor APOBEC-Cas-9 mediated editing. eGFP reporters for single base editing by APOBEC-Cas9...previous reporter, the Harris Lab created a panel of eGFP reporters to quantify the on-target DNA editing ...
  2. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    Published
    April 19, 2016, 2:30 p.m.
    ...remain unaware that EGFP/GFP is prone to forming noncovalent dimers. This property of EGFP can lead to significant...The Kd of EGFP is 0.11 mM [2]. Following the simple logic outlined above, you might think EGFP is unlikely..., cyan and yellow variants of EGFP equally capable of dimerizing as EGFP. As a result, CFP and YFP dimers...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 ...using a tag with a serious drawback. The standard EGFP plasmid that used to be sold by Clontech and is ... in the world, is not inert. In all seriousness, EGFP/GFP has a real nontrivial propensity to noncovalently...
  3. New and Upcoming Viral Vectors - September 2019

    Type
    Blog Post
    Published
    Sept. 12, 2019, 1 p.m.
    ...our entire AAV inventory. Our new AAVs include: EGFP-expressing AAV for serotype testing Calcium sensors... testing AAV, which are small (20 ul) samples of EGFP-expressing AAV packaged in various serotypes. This...complete! pAAV-CAG-GFP (plasmid 37825) and pAAV-hSyn-EGFP (plasmid 50465) are now available as 20 ul aliquots...recently released these controls: pAAV-hSyn-DIO-EGFP (50457-AAV5): The Synapsin promoter directs broad...broad, neuronal expression. AAV pCAG-FLEX-EGFP-WPRE (51502-AAV5): The CAG promoter directs ubiquitous expression...interneurons. pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP (112677-AAV5 and 112677-AAVrg): The EF1a promoter...mCherry in the absence of Cre, and expresses nuclear EGFP in the presence of Cre. Biosensor AAV (calcium ...
  4. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    Published
    May 15, 2014, 3:33 p.m.
    ...Find FRET plasmids here. Split EGFP: An alternative to FRET, split EGFP has also been used to study protein-protein...Fluorescent Protein Mutations Relative to Wildtype GFP EGFP F64L; S65T EYFP S65G; V68L; S72A; T203Y mYFP...protein-protein interactions. In this case, two portions of EGFP are fused to the proteins of interest, and when ...they come into close proximity, the two halves of EGFP undergo folding, maturation, and fluorescence. Biosensors... egfp...
  5. Plasmids 101: How to Name Your Plasmid in 3 Easy Steps

    Type
    Blog Post
    Published
    Feb. 13, 2014, 1 p.m.
    ...pBACKBONE-Flag-hGene(Q295A)-EGFP  Wildtype version: pBACKBONE-Flag-hGene-EGFP or pBACKBONE-Flag-hGene(wt)-EGFP These simple...first. pBACKBONE-Flag-hGene If there was also an EGFP fused to the C-terminal of your insert you would...would list it after the insert. pBACKBONE-Flag-hGene-EGFP Other considerations  If your insert contains a ...
  6. New and Upcoming Viral Vectors - December 2019

    Type
    Blog Post
    Published
    Dec. 19, 2019, 2:40 p.m.
    ...Nuc-flox(mCherry)-EGFP 27056 AAVrg pAAV-Ef1a-DIO EYFP 50457 AAV2 pAAV-hSyn-DIO-EGFP   Biosensor AAV...pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP 50457 AAV2 pAAV-hSyn-DIO-EGFP Recombinases Plasmid Serotype... Plasmid Serotype Name 51502 AAV5 pCAG-FLEX-EGFP-WPRE 114471 AAV1, AAV5 pAAV-Ef1a-fDIO mCherry ...
  7. New and Upcoming Viral Vectors - June 2019

    Type
    Blog Post
    Published
    June 18, 2019, 1:15 p.m.
    ... Plasmid Serotype Name 105530  AAV1 pAAV.CMV.PI.EGFP.WPRE.bGH 37825  AAV1 pAAV-CAG-GFP 59462  AAV2...50465 AAV8, 9 (20 µL size serotype testing) AAV-hSyn-EGFP Biosensor AAV The ability to detect the neurotransmitters...Name 119741  AAV5  AAV SYN flex PSAM4 GlyR IRES EGFP Recombinase AAV Finally, we are excited to add...2, 5, rg (20 µL size serotype testing) pAAV-hSyn-EGFP Recombinases Plasmid Serotype Name 55634...Serotype Name 119472  AAV5  AV SYN PSAM4 GlyR IRES EGFP   Biosensors Plasmid Serotype Name 105322...
  8. Fluorescent Protein Travel Awards - Protein Variants, a Serotonin Sensor, and an Artificial Leaf Replica System

    Type
    Blog Post
    Published
    June 4, 2019, 1:02 p.m.
    ...variant library is monitored by fusing the variants to eGFP followed by an internal ribosomal entry site and...a variant has wild-type levels of abundance, the eGFP will be stable and fluoresce. But if the variant...unstable, it will be degraded, leading to reduced eGFP fluorescence. Cells expressing the library are then...fluorescence-activated cell sorting into four bins based on their eGFP:mCherry ratios. Genomic DNA from each bin are then extracted...
  9. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    Published
    May 20, 2014, 2:06 p.m.
    .... Some proteins are reversibly switchable (e.g. rsEGFP, Dreiklang). Fluorescent Timers (FT): These proteins... by the temperature. For instance, enhanced GFP (EGFP) was optimized for 37°C, and is therefore most suited...many cases the brightness is compared to that of EGFP which is set as 1. Some proteins are very dim (e.g...Fluorescence Spectrum Viewer  from BD bioscience. ilovegfp – a site with very comprehensive data sheets on...
  10. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ...along with EGFP while retaining an emission spectrum that is easily separable from that of EGFP. This makes...spectra (N- and C-terminal tags for mTagBFP, TagRFPt, EGFP, mVenus, mCerulean3, mKOFP2) and empty vectors that...and combined it with a variety of reporters (i.e. eGFP) and effectors (i.e. DREADDs). The team demonstrated...mCyFP1, the Lin and Yasuda groups use both mCyFP1 and EGFP as FRET donors in experiments simultaneously measuring...
  11. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    Published
    June 24, 2020, 5:45 p.m.
    ...) Vlatka Zoldoš’ lab has deposited pdCas9-DNMT3A-EGFP and pdCas9-DNMT3A-PuroR for targeted cytosine methylation...methylation in mammalian cells. Co-expression markers EGFP and PuroR enable sorting and selection of transduced...Goodell's lab has deposited pcDNA3.1-dCas9-MQ1(Q147L)-EGFP, a fusion of dCas9 to a small DNA methyltransferase...epigenome-editing tools. pLV hUbC-dCas9-MQ1(Q147L)-EGFP is also available for lentiviral transduction. Lysine-specific...
  12. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    Published
    Aug. 23, 2018, 12:05 p.m.
    ...necessary to generate the strictly monomeric EGFP variant, mEGFP (Zacharias et al., 2002). GFP needs oxygen...variants. Here I use the name ‘GFP’, which refers to mEGFP. This variant was made by introducing two mutations...original jellyfish GFP (AvGFP) to create enhanced GFP (EGFP), a variant with improved brightness (Tsien, 1998...
  13. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    Published
    March 14, 2017, 2:30 p.m.
    ...Over-production of vascular endothelial growth factors (VEGF) in tumors is attributed to a signaling cascade,...factor (HIF-1), which promotes the production of VEGF to increase angiogenesis around the tumor. This ...monoclonal antibodies targeting the PD-L1 pathway, VEGF production, and CTLA4 and CD protein blocking. Other...targets under research include TIM-3, LAG-3, STAT-3, EGFR, CD20 and CD73 on specific solid as well as blood-based...
  14. New Viral Vectors - March 2024

    Type
    Blog Post
    Published
    March 5, 2024, 2:15 p.m.
    ...vector pENN.AAV.EF1a.eGFP.WPRE.rBG AAV PHP.eB Controls Wilson New serotype pAAV-CaMKIIa-EGFP AAV PHP.eB...Chemogenetics Deisseroth New viral vector pAAV.CMV.PI.EGFP.WPRE.bGH  AAV PHP.eB Controls Wilson New viral ...
  15. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    Published
    April 25, 2019, 3:01 p.m.
    ... to LanYFP and the addition of the enhanced GFP (EGFP)-type termini. mNeonGreen was reported as the brightest...protein. In vitro, mWasabi is 1.6-fold brighter than EGFP and is photostable. It also has narrow excitation...it does not significantly excite at 400 nm while EGFP does. Find mWasabi plasmids here! One of the most...
  16. New and Upcoming Viral Vectors - Spring 2019

    Type
    Blog Post
    Published
    March 26, 2019, 1:40 p.m.
    ...IRES EGFP Chimeric channels for neuronal manipulation 119742  AAV5  AAV SYN PSAM4 GlyR IRES EGFP Chimeric...*  pAAV-CAG-GFP 50465  AAV8*, AAV9*  pAAV-hSyn-EGFP *Also coming in our new 20 ul trial size! Voltage...
  17. Choosing Your Fluorescent Proteins for Multi-Color Imaging

    Type
    Blog Post
    Published
    Oct. 9, 2014, 3 p.m.
    ...proteins for imaging with this set are mTagBFP2, EGFP or one of the improved GFP variants, mRuby2 or TagRFP-T...mammalian cells, one of the improved folding variants of EGFP like mEmerald or Clover is probably best; mNeonGreen...and have reported brightness measurements. Here, EGFP outperforms the improved folding variants, presumably...
  18. New and Upcoming Viral Vectors - May 2020

    Type
    Blog Post
    Published
    May 28, 2020, 1:15 p.m.
    ...pAAV EF1a Nuc-flox(mCherry)-EGFP Brandon Harvey 50457 AAV2 pAAV-hSyn-DIO-EGFP Bryan Roth 50459 AAV1 pAAV-hSyn-DIO-mCherry...pAAV-hSyn-DIO-mCherry Bryan Roth 50457 AAV1 pAAV-hSyn-DIO-EGFP Bryan Roth 114472 AAVrg pAAV-hSyn-mCherry Karl...
  19. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    Published
    Sept. 22, 2015, 2:30 p.m.
    ...receptor (EGFR) with the ClonTracer library and monitored population dynamics after treatment with EGFR inhibitor... in a study focusing on tumor cell resistance to EGFR inhibitor (Hata et al., 2016).  Bhang deposited ...
  20. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    Published
    June 28, 2016, 10:59 a.m.
    ...halves of EGFP to recombine by homology directed repair, and resulting in the expression of EGFP. Using ...indicates where your target is inserted, disrupting the EGFP fluorescent signal, for details on this plasmid,...
Showing: 1 - 20 of 59 results