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We narrowed to 527 results for: HA tag

Showing: 161 - 180 of 527 results

  1. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ... Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs... can be used in cells or organisms in which Cas9 has been integrated into the genome. gRNA Empty Vectors...Marraffini pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) 47868 Mammalian U6 yes, cut N. meningitidis...pyogenes tagRFP Ebert pL-CRISPR.SFFV.tRFP 57826 Mammalian/Lentiviral BsmBI yes, cut S. pyogenes tagRFP Ebert...Mammalian/Lentiviral BbsI none S. pyogenes Puro, TagBFP Yusa lentiGuide-Puro 52963 Mammalian/Lentiviral...Mammalian/Lentiviral BsmBI yes, cut S. pyogenes tagRFP Ebert pFGC-pcoCas9 52256 Plant AscI, PacI, SbfI...57824 Mammalian/Lentiviral BsmBI none S. pyogenes tagRFP657 Ebert pL-CRISPR.SFFV.GFP 57827 Mammalian/Lentiviral...

  2. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...CRISPR plasmids at Addgene! Knock-in: Introducing tags or stop signals Once Cas9 generates double-strand...incorporating very long ncRNAs is impractical. Instead, tags or stop elements can be inserted to disrupt the ...CRISPR systems for targeting ncRNAs While CRISPR/Cas9 has proven to be an extremely useful system for modifying...This system is often used to knock down lncRNAs and has fewer off-target effects. Newer Cas protein variants...transcriptional activators such as VP64, SAM, or SunTag to specific ncRNA loci to enhance or upregulate...

  3. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...short sequences of up to 100 bp, such as epitope tags, design primers with template binding regions to...combining multiple primers, a C-terminal epitope tag can be moved to the N-terminus of a protein-coding... can be introduced in a gene, while adding a GFP-tag (Figure 3). Achieving multiple modifications requires..., TOP10, and Mach1. No common lab cloning strain has failed so far. What is most important is the transformation... to the 5’ end of the reverse primer. Mutagenesis - Mutagenesis is performed by designing primers astride...modification, from insertions, deletions, and point mutagenesis to complex assembly of multiple genes. Primer...rapid method for recombination and site-specific mutagenesis by placing homologous ends on DNA using polymerase...

  4. Antibodies 101: The Four ELISAs and When to Use Them

    Type
    Blog Post
    ...The antibodies are conjugated with a reporter or tag that can then be detected.  Typically, the reporter... (HRP) or alkaline phosphatase (AP). Fluorescent tags may also be used, but for simplicity we will discuss...testing a recombinant protein, especially one with tags, be sure to include a sample that expresses the ...While the method is simple and saves time, it also has several drawbacks. Direct ELISAs generally have higher... of the advantages and disadvantages of each ELISA method Method Advantages Disadvantages Direct...rundown of the different types of ELISA, advantages and disadvantages of each, and some considerations when...isotype.     Specificity and timing are the major disadvantages of the indirect method. The assay only uses ...

  5. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...and gram negative bacteria periplasm. Efforts to tag endogenous genes for proteins in these compartments...Munro, Sean, and H. R. Pelham. "Use of peptide tagging to detect proteins expressed from cloned genes:...Avaliable at Addgene Read Our Blog Post on Protein Tags Read Other Posts on Fluorescent Proteins Browse ...enabled numerous cell biology assays. However, there has been a pervasive view that most FPs in a cell will...FP-fusion proteins functionally relative to the untagged protein of interest. To avoid the accumulation...

  6. Mapping the 4D nucleome with CRISPR/Cas9

    Type
    Blog Post
    ... from this publication at Addgene. A protein-tagging system for signal amplification in gene expression...nuclease activity, catalytically dead Cas9 (dCas9) has many applications, the most well-known being transcriptional...transcriptional activation/repression. dCas9 has also previously been adapted to fluorescently label a sequence...imaging time for this technique. The NIH Common Fund has made mapping the 4D nucleome a specific priority,...ability to label live cells make this system truly advantageous. This new technique, developed in Thoru Pederson...sensitivity of the technique. One potential solution is SunTag, a synthetic scaffold that can be used to recruit...from Addgene!) Since the signal level provided by SunTag is so high, cells may be imaged under lower illumination...

  8. New and Upcoming Viral Vectors - Spring 2019

    Type
    Blog Post
    ...AAV5  pOTTC1596 - pAAV SYN1 HA-hM3D(Gq) 121538  AAV5  pOTTC1484 - pAAV SYN1 HA-hM4D(Gi) 119741  AAV5 ... vectors for experiments encoding a fluorescent-tagged molecular tool.   Find pAAV-FLEX-tdTomato, pAAV-GFAP104...iGABASnFR GABA sensor (pAAV.hSyn-FLEX.iGABASnFR) has good membrane localization and brightness. The F102G...its F102G variant in AAV1 at Addgene! Archon1, a voltage sensor AAV Historically, electrical activity in...measured with implanted electrodes. The Archon1 voltage reporter allows a way to monitor neuronal electrical...Archon1-KGC-GFP-ER2]) of this membrane-embedding voltage detector. Find Archon1 and its cre-dependent version...pAAV-hSyn-EGFP *Also coming in our new 20 ul trial size! Voltage indicators Plasmid Serotype Name 119036  AAV1...

  9. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ...pair changes, and the addition of protein tags. It also has the flexibility to modify the E. coli chromosome...non-selectable DNA fragments, such as gene replacements and tags.  The typical frequency of recombinants is 1 positive...recombineering system The lambda red recombineering system has three components (Figure 1): 1) Exo, 2) Beta, and... in the center of the sequence. Since lambda red has a higher recombination frequency when the lagging...activating MMR: 1) use a strain of bacteria that has key MMR proteins knocked out or 2) specially design...recombinant clones If a antibiotic resistance gene has been inserted, recombinants can first be selected...repress expression of exo, beta and gam.  A major advantage of using this method for lambda red expression...

  10. Lentivirus Plasmids

    Type
    Collection
    ...randomly into the host genome, it has the potential for insertional mutagensis. The 2nd and 3rd generation ...

  11. Optogenetics Guide

    Type
    Guide
    ...function. TULIPs Tunable light-inducible dimerization tags (TULIPs), plasmids allow cloning of protein coding...sequences with GFP-LOVpep, cpPDZ, ePDZb and ePDZb1 as tags. pDawn/pDusk Blue-light photoreceptor for light-...turning on and off the light source. Optogenetics has been broadly applied to study the physiology of the...channel. ChR2 is preferred over ChR1 because ChR2 has higher conductance at physiological pH and trafficks...Depolarizing) ChR Variants. The optogenetic toolbox has been expanded by scientists through both the identification...heterodimerization can reconstitute the protein. This principle has also permitted the design of synthetic two-part transcription...Phytochrome, cryptochrome, and LOV (light oxygen voltage)-based systems have been used in many experimental...

  12. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ...Plasmids Find Empty Backbones with Fluorescent Protein Tags  ... first publication in 2012, the Nagai laboratory has assembled a collection of multicolored nano-lanterns... low luminescence By itself, Renilla luciferase has a low quantum yield, and when transiently expressed...using chemiluminescent proteins for imaging, but it has precluded widespread adoption of the technology. ...Nano-lanterns and your experiments The Nagai laboratory has a done a wonderful job characterizing Nano-lanterns...described the first Nano-lantern, a fusion of a mutagenized Renilla luciferase (RLuc8) with Venus. This luminescent...

  13. New and Upcoming Viral Vectors - June 2019

    Type
    Blog Post
    ...AAV2 pAAV-CaMKIIa-HA-hM4D(Gi)-IRES-mCitrine 121538  AAV5 pOTTC1484 - pAAV SYN1 HA-hM4D(Gi) 121539 ...for activating and inhibiting neurons. Minimally-tagged DREADD AAV from the Richie lab are especially useful...121539  AAV5 pOTTC1596 - pAAV SYN1 HA-hM3D(Gq) New chemogenetics AAV Plasmid Serotype Name 119741  ... AAV9 pAAV-hSynapsin1-FLEx-axon-GCaMP6s New voltage sensors Plasmid Serotype Name 119036  AAV1...

  14. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

    Type
    Blog Post
    ...fusion protein consisting of sfGFP fused to a His-tagged ABC transporter known to bind to the HMO of interest... dronpa fusions. With these various designs, Xin has been able to develop photoswitchable Intersectin,...Intersectin, Cdc42, Raf1, Mek1, Mek2, Cdk5, and Cas9. Xin has demonstrated the utility of these FLiPs in a variety... of cells that actively produce the HMOs. Fatima has demonstrated the successful production of both types...precisely spatio-temporally controlled as a key advantage over other inducible enzyme technologies. She ...

  15. Another Pathway into Cells: iTOP

    Type
    Blog Post
    ...surprising discovery that both purified CPP-tagged protein and an untagged control could enter cultured cells ...derived from the same cell type; however, their usage has been limited by technical problems. For instance,...binary systems in which transient protein expression has a long-lasting effect, such as Cre- or Cas9-mediated...effects. Third, protein purification (once a protocol has been established) may be less laborious than viral...used for protein purification must be helping the untagged protein enter cells. Subsequent work found that...stem cells, they found very high recombination percentages (78-79% after two rounds of transduction.) Compared...

  16. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...cells according to whether or not they have been tagged with a fluorescent protein or dye. It separates... distinguishing cell populations that have been tagged with different fluorescent proteins or dyes. 3.... cell mobility within these complex environments has never been easy and, for many years, researchers ...Intravital imaging powered by two-photon microscopy has uncovered host-pathogen interactions leading to the...proteins can be used in your experiments, experience has shown that some are more reliable and more efficient...important to finely tune the excitation wavelength as it has been shown that even a small incremental change of...this system called Brainbow, the Ubow mouse strain has been created to fate map Langerhans cells and follicular...

  17. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    ...into a vector with a large tag at the N-terminus, but you don’t have the tag already included in the vector...you can insert both the gene of interest and the tag sequences into the vector in one step without scars...where the vector that you want to use is very large, has a high GC content, contains a lot of repeats--any...In this blog post, I will go over some advantages, disadvantages, and examples of how scientists are using... common, and for good reason. They offer many advantages over the traditional restriction enzyme cloning...thereof) nor a serious time commitment. Another advantage is that this process makes it easy to generate...will be using. Some of these products may offer advantages that Gibson assembly does not such as increased...

  18. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ... be selected, since it cleaves at the stop codon TAG. Design your repair template with 55-100 base...mScarlet).   Plasmid ID Plasmid Name Tags 69090 pMJ915 MBP (N terminal); 6xHis (N ...CRISPR genome editing has made it easier to create knockout alleles in a variety of species, including...including creating Cre-dependent conditional alleles, has remained a challenge. Enter Easi-CRISPR: a method...left homology arm guanine (i.e. GG); this design has been shown to increase T7 promoter activity. To avoid...

  19. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    ...respectively. mNeonGreen can be used as a fusion tag for a wide range of applications from traditional...single-molecule super resolution imaging. mNeonGreen also has little sequence identity to other common fluorescent...fold brighter than EGFP and is photostable. It also has narrow excitation and emission peaks (max excitation...was then screened through successive rounds of mutagenesis for the brightest teal fluorescent proteins that...scientists from Florida State University, they used a mutagenesis approach and found that specific amino acid changes...

  20. Cre-Lox and Other Site-Specific Recombinases

    Type
    Collection
    ...genomic attP site. So far, their practical application has been limited by low integration efficiency but research...(Length) Common Variants Cre loxP ATAACTTCGTATAgcatacatTATACGAAGTTAT 13 bp inverted repeats + 8 bp spacer...site is widely used. Flp FRT GAAGTTCCTATTCCGAAGTTCCTATTCtctagaaaGTATAGGAACTTC 13 bp inverted repeats + 8... F3, F5, FRT14, FRT15 minimal FRT GAAGTTCCTATTCtctagaaaGTATAGGAACTTC Table 1. Summary of Cre, Dre, and...turning on another. The FLEx switch system takes advantage of the orientation specificity of Cre and the ...
Showing: 161 - 180 of 527 results