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We narrowed to 46 results for: Free Use GFP

Showing: 1 - 20 of 46 results

  1. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    Published
    Sept. 21, 2021, 1:15 p.m.
    ...find and use the plasmids you need. Here's what you'll find in this article: An improved cell-free system... The CRISPR Corner New ready-to-use viral preps   Improved cell-free transcription/translation system...successfully reconstituted in this cell-free system. In summary, this cell free system allows for a relatively ...vectors and flow cytometry is used to sort populations by mCherry and GFP expression to check for ABE’s...examine biosynthetic pathways  By Gabrielle Clouse Recently, cell-free transcription/translation systems (also...that produce useful natural materials, such as antibiotics. Simon Moore and Paul Freemont’s lab previously...Graphical Abstract: Schematic representation of cell-free system, capable of producing fluorescent proteins...

  2. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ...Gagoski used SITS to create the pCell-Free vectors, Gateway-compatible backbones that enable cell-free expression...can be used, and how they’ve been used to date . If you have any questions, please write to us in the ...N-terminal eGFP tag carried on all constructs. The Cell-Free Protein Expression Kit has been used to produced...sequences used in this so-called bicistronic design (BCD). The 49 vectors in this kit can be used to drive...protein of interest. The plasmids used to generate these fusions all use the same restriction enzymes for...Purification Process with pCri Cell-free protein expression kit Cell-free expression systems have been improved...found at the 3’ end of the leader sequence. Using a variety of GFP and RFP fusions as their test genes, the...

  3. Hot Plasmids - May 2022

    Type
    Blog Post
    Published
    May 26, 2022, 1:15 p.m.
    ...2021.    BLITZ uses the biotin ligase TurboID fused to a conditionally stabilized GFP-nanobody rather...line expressing a GFP-tagged version of a protein of interest. The binding of the GFP nanobody to its target... proximity to the GFP-tagged protein (albeit slightly offset due to the nanobody-GFP interaction). The...be deconvoluted using freeware.    Fig. 4: Images of 12 visual barcodes in use in A375 cell line...Fig. 3: Development of a cleavage-free dCas9-based gene editor using microbial SSAPs. Image from Wang,... hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like to write...recently created a flexible system that makes use of pre-existing transgenic lines, termed BLITZ (Biotin...

  4. Our Most Popular Blog Posts from 2019

    Type
    Blog Post
    Published
    Dec. 31, 2019, 2:26 p.m.
    ...Toolbox: Free Tools for Making Scientific Graphics by Beth Kenkel Find a round-up of tools you can use for...scientific graphics. PS: they’re free. 3. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right...you’d like to see more of? Let us know by leaving a comment or emailing us at [email protected].  Thank you...toxin-antitoxin systems and sacB counter-selection) tools used in cloning. 8. Bright Monomeric Fluorescent Proteins...people are using for their research. In this blog post, we’ve covered five model organisms: mouse, fruit ...Joachim Goedhart shares some strategies for generating GFP fusion proteins. From considering protein size and... some of these top blog posts. We’d also like to use this time to hear some feedback from the community...

  5. 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...Endotoxin contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize...addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.     Figure...titers obtained using ddPCR are generally two to three fold higher than those achieved using the standard...AAV preps and our lentiviral preps.   Viruses are generated using standard methods that have been optimized...of unexpected DNA contaminants. The data is also used to determine recombination rates in FLEX/DIO/fDIO...protein present in the sample is also determined and used to determine purity of the AAV preparation. ...

  6. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ... the GFP-Puro will be inserted just upstream of a stop codon. One potential concern is if the GFP-Puro...Integration into Target Chromosomes). Using their PITCh plasmids, GFP knock-in cell lines can be created ...template with flanking sequence homology for error-free DSB repair. NHEJ joins the ends of a DSB in an error-prone...insertions and deletions common. In contrast, MMEJ uses regions with 5-25 bp of microhomology flanking a...inefficiency of HDR for knock-ins, some labs have used NHEJ for whole plasmid integration; however, since...detailed protocol for MMEJ-mediated knock-in of a GFP-Puro cassette into a given locus, just upstream of...microhomology to the insertion locus flanking the GFP-Puro cassette. Three double stranded breaks are necessary...

  7. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...antibodies against them, fused them to GFP, and compared them to popularly used epitopes tags such as HA...to complement the use of optogenetic tools, where light used for excitation can cause unintended activation...both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins...plasmids for use in high-throughput screening of pre-mRNA splicing. This reporter system can be used for identifying... that can be used for monitoring cellular dynamics. These fusion proteins have been used as subcellular...Compartment mCherry-Rab5 BFP-Rab5 GFP-Rab5B Vacuolar & Budding Compartment GFP-Rab4B Mitochondria mito-BFP...GTPase in division) GFP-Mff (outer membrane protein) Late Endosome mCherry-Rab7A GFP-Rab7A     ...

  8. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

    Type
    Blog Post
    Published
    June 27, 2018, 1:16 p.m.
    ...nanobodies described here can be used to target any protein fused with GFP, YFP, mCerulean, or other related...for quantitative analysis using different detection methods. Since the anti-GFP nanobody is able to bind...a set of plasmids each containing an anti-GFP nanobody fused with a different functional element (tyrosine...University of Massachusetts Medical School have developed a group of plasmids (here) that can be used to label... hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like to write...facilitating techniques that are dependent on long, error-free, reverse transcription of RNA such as long-read ...that associate with specific genomic loci. He made use of catalytically inactive dCas9 by fusing it to an...

  9. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    Published
    March 7, 2017, 3:30 p.m.
    ...editing. A virus encoding GFP plus a gRNA targeting GFP (pXPR_011, 59702-LV) was used to assay Cas9 activity...from BRAF-edited and GFP-edited amplification products (BRAF-GFP duplex) and from GFP-edited amplification... 1. Use multiple gRNAs to target different regions of your gene. I used existing BRAF gRNA viruses, but...I thought I could see GFP expression with a fluorescence microscope, but the GFP expression was so low...expressed GFP) that I couldn’t get a good image. You could probably also do Western blotting against GFP expression...76480-LV) or control GFP gRNA (BRDN0000561167) (80034-LV) were individually used to transduced stable ...product indicates a gene edit. GFP gRNA-transduced Cas9-A549 cells are used as a control. DNA was analyzed...

  10. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    Published
    Sept. 6, 2022, 1:15 p.m.
    ...literature, scientists can use the Data Hub to compare different methods and vectors, and use this data to guide...when others have already answered this question and used it to publish!  Working with AAV vectors in neuroscience... I started working with AAV vectors. I wanted to use an AAV vector to express ChR2(H134R) in A2 norepinephrine... cassette with an ubiquitous promoter to express GFP in a group of young rats. We found that, in this ...important factors such as what is the best promoter to use, or if and how much the spread of the virus to adjacent...label norepinephrine (NE) expressing neurons and GFP (green) to label neurons transduced by each serotype... (3,5,8), injection titers (1,2), and cell types used for packaging viral particles (6), can influence...

  11. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ...capture module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph of H1.8-GFP nucleosomes collected...many future variations using nanobodies or scFvs for other targets or could be used for applications beyond...that this method of delivery could be used to partially correct mouse models of genetic blindness (An et ...epigenetic silencing, which can be toxic, CHARM uses a cofactor fused to a histone tail to recruit and stimulate...In MagIC-cryo-EM, target proteins (expressed as a GFP fusion in this proof-of-concept study) are captured...SpyTag-SpyCatcher linkers and spacer proteins ending with a GFP nanobody (Figure 1). This enrichment streamlines ...) 3D model of the obtained structure of the H1.8-GFP-bound nucleosome. Images adapted from Arimura et ...

  12. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    Published
    April 17, 2018, 1:37 p.m.
    ...minicircle RNA Delivery DNA-Free High High Transgene-free and vector-free Multiple transfections required...required Protein Delivery DNA-Free Low High Transgene-free and vector-free. Slow and inefficient Difficult...Excisible Medium Medium Generates transgene-free and vector-free cells Genome integration Need to sequence...Non-integrating Low Medium Generates transgene-free and vector-free cells No genomic integration Inefficient...Non-integrating Medium Medium Generates transgene-free and vector-free cells No genomic integration Can be difficult...usually Medium Medium Generates transgene-free and vector-free cells Limited genomic integration Multiple...Non-integrating Medium Medium Generates transgene-free and vector-free cells No genomic integration Inefficient...

  13. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    Published
    May 15, 2018, 12:43 p.m.
    ...researcher can cause them to interact by stimulation with blue light.  Similarly, light can also be used to control... hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like to write...Institute for Basic Science (IBS) has developed another useful optogenetic tool that enables robust and efficient...without the extra peptide. CRY2clust should prove a useful tool to further probe cellular function. The CRY2clust...segment Optogenetic stimulation is a powerful tool used by neuroscientists to interrogate neuronal networks...plasmids described in this work including the soCoChR-GFP plasmid as well as several derivatives encoding alternative...podcast segment With the help of engineered microbes, used coffee grounds could be economically repurposed ...

  14. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    Published
    June 3, 2021, 1:15 p.m.
    ... are scFvs used for? scFvs are used in many of the ways antibodies are. scFvs are even used to screen ...GCN4 binding scFv that’s fused to GFP. When the multiple copies of the scFv-GFP fusion bind the SunTag ...clearance, which is useful for imaging applications; better tissue penetration which is useful for therapeutic...immunized animals, but it's possible to avoid animal use by using semi-synthetic or entirely synthetic VL and ...contains an HA-binding scFv that’s fused to a fluorescent protein such as GFP. The HA frankenbody works just... this, one or more of these approaches are often used for scFv expression in bacteria:  targeting to ...Sandomenico et al., 2020). conversion to a cysteine free format that’s easier to express in the cytoplasm...

  15. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    Published
    Aug. 23, 2018, 12:05 p.m.
    ... There are numerous GFP variants. Here I use the name ‘GFP’, which refers to mEGFP. This variant was made...2013; Hilaire et al., 2017). GFP can only be used for tagging proteins GFP fusion proteins are made by ...S65T) in the original jellyfish GFP (AvGFP) to create enhanced GFP (EGFP), a variant with improved brightness...Microscopy (University of Amsterdam). GFP is the most popular, most widely used genetically encoded fluorescent...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... a fusion with GFP allows for detection of genomic locations (Chen et al., 2013). GFP fluorescence overlaps...

  16. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    Published
    July 25, 2019, 12:03 p.m.
    ... We used the disarmed Agrobacterium tumefaciens strain GV3101(pMP90), which works well for us. Use a well...Fluorescent proteins (FP, e.g. GFP) can be used in many ways. Often a FP is fused to a protein of interest ...can be used to develop stable hairy root lines in tissue culture. This method is commonly used in C. roseus...the commonly used CaMV 35S promoter in Agrobacterium. This can be easily avoided by the use of introns,...gene silencing (VIGS) is a transient technique that uses a virus and the plant's post-transcriptional machinery...Mortensen et al., 2019). Our method might provide some useful guidelines for the development of transient transformation... If you are new to the field, the following are useful tips for your first transient leaf transformation...

  17. Plasmids 101: Protein tags

    Type
    Blog Post
    Published
    Dec. 11, 2014, 4:26 p.m.
    ...Plasmids 101 has covered GFP and its related fluorescent proteins, which are sometimes used as tags for detection...Affinity Frequently used for western blots, IP, co-IP, IF, flow -cytometry, but rarely used for purification...Fluorescent Protein Should I Use? Plasmids 101: Green Fluorescent Protein (GFP) Fluorescent Protein Guide...various mutations used to increase its stability activity, can be readily purified using plasmids found ... proteins can be purified using metal-affinity chromatography, usually using a Ni2+ column. Like other...recognition site. In fact, SUMO cannot be used in eukaryotes because there is already too much SUMO protease...the accompanying cartoon, they have a multitude of uses including (but not limited to) purification, detection...

  18. 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...encoded tetracysteine (Cys4) peptide tag could be used to label proteins in cells (Figure 1a). Other strategies...e.g., SNAP-tag, HaloTag, TMP-tag) – These widely used systems consist of a genetically encoded enzyme ...be incorporated into a protein structure and then used with the growing toolbox of bioorthogonal chemical...meaning the ligand exhibits low fluorescence when free in solution, but high fluorescence upon binding ...polar groups (Figure 2b), which can preclude their use inside living cells. We recently discovered that ...

  19. 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.
    ...simplest and best solution is to use truly monomeric FPs. EGFP and other GFP family members (superfolder, ...contributed by guest blogger Erik L. Snapp. Stop using EGFP/GFP for fusion proteins! Despite multiple studies... If you're using green fluorescent protein or Enhanced Green Fluorescent Protein (GFP/EGFP) for a transcriptional...between, you are using a tag with a serious drawback. The standard EGFP plasmid that used to be sold by ...remain unaware that EGFP/GFP is prone to forming noncovalent dimers. This property of EGFP can lead to significant...in a freezer box in just about every lab in the world, is not inert. In all seriousness, EGFP/GFP has ...designated mEGFP, mVenus, etc. A word of caution is needed because FPs not derived from GFP, i.e. the ...

  20. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    Published
    Dec. 7, 2021, 2:15 p.m.
    ...which can be used for most antibodies compatible with either Protein A or G.  While commonly used, Protein...the antibody used for IP is derived from the same species as the antibody that will be used for detection...such as this, an epitope such as c-Myc, GFP or V5 can be used to tag the protein of interest on the C-...will need to be verified using alternative methods.  Finally, IP can also be used to study proteins that...Immunoprecipitation (IP) uses immobilized antibodies, or immunoglobulins, to isolate a specific protein...protein out of a complex mix. Using this technique, users can look for the presence or absence of a protein...target is present in low amounts. In this method, the free antibody is incubated with the protein mix and antibody-target...
Showing: 1 - 20 of 46 results