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We narrowed to 163 results for: NAP

Showing: 1 - 40 of 163 results

  1. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    ...powered by GSL Biotech's SnapGene Server Software. With the backing of SnapGene’s sequence viewer software... Overview of SnapGene powered plasmid maps at Addgene  Three factors make our SnapGene powered plasmid...the features you need quickly. SnapGene’s Feature Library - SnapGene’s software seamlessly identifies ...example below. Our old map is on the left while the SnapGene powered map is on the right (click here to see...t detected by our in-house software. In fact, SnapGene’s feature library includes nearly 3x as many features...identify. Improved Restriction Enzyme Handling - SnapGene’s software positions restriction enzyme annotations...you can download an annotated GenBank File or a SnapGene file for the sequences listed. You can then analyze...

  3. Identifying Sequence Elements with SnapGene's Feature Database

    Type
    Blog Post
    ... who uses SnapGene or the free SnapGene Viewer companion product. If you don’t use SnapGene yet, you can...guest bloggers Aline and Benjamin Glick from SnapGene. SnapGene was created to meet a need. While there were...approach is SnapGene’s algorithm for detecting common features. This algorithm enables one of SnapGene’s most... us at snapgene.com. Many thanks to our guest bloggers Aline and Benjamin Glick from SnapGene!  Ben Glick...Addgene Blog Read about our SnapGene powered plasmid maps Learn how we use SnapGene during the plasmid quality...properties of the DNA molecules they were using. SnapGene was created to alleviate these problems through...make specialized controls available when needed. SnapGene has been engineered to be easy and enjoyable to...

  4. Plasmids 101: Walkthrough of Addgene’s Snapgene-Powered Quality Control Process

    Type
    Blog Post
    ...through our Snapgene powered maps and sequence analysis tools.     The main ways we use SnapGene for plasmid...version of Snapgene to visualize the sequence and look for features. The first thing that the Snapgene software...proteins that are already included in Snapgene’s extensive database. Snapgene also provides easy visualization...software as the desktop version of Snapgene. When you download a Snapgene file directly from our website,...website, you can open it with either the Snapgene viewer or Snapgene Desktop programs to analyze the sequence... plasmids. We’ve also talked about how our new Snapgene generated maps provide improved feature detection...with an easy to use interface. We regularly use Snapgene for our quality control process because of its...

  6. Immunology Research Plasmids and Resources

    Type
    Collection
    ...CXCL8, GCP-1, GCP1, LECT, LUCT, LYNAP, MDNCF, MONAP, NAF, NAP-1, NAP1 IL8RA interleukin 8 receptor, alpha... activity, alpha) FSP, GRO1, GROa, MGSA, MGSA-a, NAP-3, SCYB1 CXCL10 chemokine (C-X-C motif) ligand 10...-III, CTAP3, CTAPIII, CXCL7, LA-PF4, LDGF, MDGF, NAP-2, PBP, SCYB7, TC1, TC2, TGB, TGB1, THBGB, THBGB1...

  9. Lentivirus Production

    Type
    Protocol
    ...°C for several hours but should be aliquoted and snap frozen in liquid nitrogen and stored at -80 °C as...

  10. Rabies and Neuronal Tracing

    Type
    Blog Post
    ...transmitted to adjacent neurons via synapses (i.e., transmitted transsynaptically). This is because the recombinant...Rabies Virus: Converting a transsynaptic neuronal tracer into a monosynaptic (first order) neuronal tracer... able to replicate, and for a virus to cross a synapse from one cell to another, it must replicate. Modified...replicates within that cell, and nascent RABV cross synapses to retrogradely infect neurons that project to...higher order neural connections. While this transsynaptic spread is useful for neuronal tracing, it has...starter cell types, as well as to only spread monosynaptically from these cells. These modifications take...express EnvA unless it is supplied in trans. Monosynaptic RABV spread can be achieved by modifying the...

  11. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...retrograde labelling, synaptic restriction as well as controlled monosynaptic and polysynaptic spreading. AAV... AAV9 are capable of synaptic spread. The underlying mechanisms of trans-synaptic trafficking are not ...., and Callaway, E.M. (2007). Monosynaptic Restriction of Transsynaptic Tracing from Single, Genetically...sending information onwards to other neuronal cells. Synapses at the terminals of axons form connections to ...the prefrontal cortex, but also indirectly via synapses to neurons in other brain regions. Further, the...tracing compounds is that their diffusion across the synaptic cleft is usually severely impaired, and only few... are transported from one neuron to another at synapses resulting in faint signals (Schwab et al., 1979...

  12. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    ...became known as SNAP-tag. There is also a fast-reacting version of SNAP-tag called SNAPf. eDHFR: The Virginia...fluorescein) or ReAsH (red, based on resorufin). SNAP-tag: In 2003, Kai Johnsson’s lab developed a self-labeling...2008). Although based on the same original enzyme, SNAP-tag and CLIP-tag show good specificity for their...Biarsenical dyes (FlAsH, ReAsH) Few Easy SNAP-tag 182 Benzylguanine (BG) Many Easy ...often up to 10-fold more). So, labeling a HaloTag or SNAP-tag construct can help you better detect dim structures...magenta) and outer mitochondrial membrane labeled with SNAP-OMP25 with Atto590-BG (green), imaged with stimulated...yellow) at the tip. 649-mDia1 and 549-CP are both SNAP-tag fusions, purified and labeled separately with...

  13. Hot Plasmids: Summer 2025

    Type
    Blog Post
    ...found at excitatory synapses (Rivera & Huang, et al., 2025). AF was fused to a presynaptic vesicle protein...remain on the presynaptic cell membrane while AF, fused to a payload, is released into the synapse. Once AF...and AAV11 packaging plasmids Mapping anterograde synaptic connections with ATLAS Bright and photostable...nucleus, endosomes, lysosome, golgi, ER, inhibitory synapses, and cytoskeleton components) and in a FRET-based...packaging plasmids here!   An ATLAS for anterograde synaptic connections By Emily P. Bentley While several ...lack of genetic control. ATLAS (anterograde transsynaptic label based on antibody-like sensors) changes...AF binds to the postsynaptic AMPA receptor and is endocytosed, the Cre recombinase payload activates a...

  14. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...neurons and their synapses.   How can you label a single neuron and its synapses in the living brain...cytoplasmic markers (e.g. eYFP) and synaptic proteins (e.g. postsynaptic PSD95-mcherry) are used in high ... a single cell with all its synapses and how it rearranges these synapses in the living brain over time...mix to label neuronal morphology (eYFP) and the synaptic protein PSD95 (PSD95-mcherry) under the control... if the cellular structure of a neuron and its synapse should be visualized at the same time from two ...Expression of a morphological marker (eYFP) and synaptic marker (PSD95-mcherry), under the control of Flp...CreERT2), visualizes individual neurons and their synapses in a conditional knockout mouse. Tamoxifen injection...

  15. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    ...Cre-expressing cells hSynap: Human Synapsin-1 promoter. Good for neuronal expression. hSynap-FLEX: Synapsin-1 promoter...and accurately differentiating between synaptic vs. extrasynaptic glutamate release. Building on the previous... for expression in Cre-expressing cells. hSynap-FLP: Synapsin-1 promoter, FLP-dependent expression GFAP...in vivo. Glutamate plays a variety of roles in synaptic communication and can trigger other forms of neuronal.... iGluSnFR3: Improved Glutamate Indicators for Synaptic Imaging However, the first two generations of ...that are more sensitive and better localized for synaptic glutamate response than its predecessors. The ...activation kinetics and localiation for imaging synaptic transmission” bioRxiv (2022)...

  16. Hot Plasmids - October 2020

    Type
    Blog Post
    ... the synaptic scaffolding proteins PSD95 and gephyrin to label excitatory and inhibitory synapses, respectively...bind to the endogenous synaptic proteins allowing them to localize to the synapse and minimize extraneous...inhibitory or excitatory synapses or that are Cre-inducible, researchers can explore synaptic integration and ...post: Tools for labeling excitatory inhibitory synaptic proteins Fluorescent protein system for C. elegans... service Labeling excitatory and inhibitory synaptic proteins with FingRs By Jason Nasse Since the ...billion neurons in the human brain. But studying synaptic connections and physiology is particularly challenging...selectively visualize excitatory and inhibitory synapses in the brain.  Figure 1:Schematic diagram...

  17. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ...Figure 2: CLIP-SNAP fusion proteins for single-molecule FRET. A) Schematic of a CLIP-SNAP fusion protein...doi.org/10.1101/2024.02.11.579803).  CLIP, CLAP, SNAP! Self-labeling tags for single-molecule biophysics...Twelvetrees Lab took advantage of the self-labeling tags SNAPf and CLIPf. These tags rely on synthetic dye molecules...combining an N-terminal CLIPf tag and a C-terminal SNAPf tag; see Figure 2), the authors developed a cost-effective...KIF5B as well as backbones for generating other SNAPf-, CLIPf-, and CLAP-tagged proteins for mammalian...other proteins of interest too! Find CLIP, CLAP, and SNAP plasmids here! Smith, E.R., Turner, E.D., Abdelhamid...indicator with enhanced sensitivity for unitary synaptic events By Brian O’Neill The Lin Lab and collaborators...

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

    Type
    Blog Post
    ...our guest bloggers Erik L. Snapp and Lindsey M. Costantini! Erik Lee Snapp received his PhD in Molecular... post was contributed by guest bloggers Erik L. Snapp and Lindsey M. Costantini. "You underestimate the...have reported that FP cysteine residues form inappropriate disulfide bonds in the secretory pathway (Figure...localization. For example, the formation of inappropriate inter-chain disulfide bonds between FPs blocks...research was conducted in the laboratory of Erik Lee Snapp and focused on optimizing fluorescent proteins for...sarcoma-associated herpesvirus.       References 1. Snapp, Erik. "Design and use of fluorescent fusion proteins... 18228466. Pubmed Central PMCID: PMC2875081. 2. Snapp, Erik Lee. "Fluorescent proteins: a cell biologist's...

  19. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ...and putative pre-synaptic terminals with one vector. AAV hSyn FLEx mGFP-2A-Synaptophysin-mRuby from Liqun...enhance axonal labeling and synaptophysin-tagged mRuby to enrich pre-synaptic labeling. Now available in...cytoplasmic targeted tdTomato and pre-synaptic targeted synaptophysin tagged eGFP. AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE...ready-to-use AAV5 viral vector. Synaptophysin-tagged GCaMP6s is enriched in the pre-synaptic terminal providing better...as a ready-to-use viral prep in serotype AAV1. Synapse targeting GCaMP6s from the Rylan Larsen lab are...

  20. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...DE3)   Basic IPTG-inducible strain containing T7 RNAP (DE3) F- ompT lon hsdSB (rB- mB-) gal dcm (DE3)...Tetracycline Arabinose-inducible expression of T7 RNAP; IPTG may still be required for expression F- ompT... ompT lon hsdSB (rB- mB-) gal dcm araB::T7RNAP-tetA General protein expression  BLR (DE3) Tetracycline...DE3)* Chloramphenicol (pLemo) Rhamnose-tunable T7 RNAP expression alleviates inclusion body formation. ...  T7 Express   IPTG-inducible expression of T7 RNAP from the genome; does not restrict methylated DNA...system. The DE3 lysogen expresses T7 RNA polymerase (RNAP) from the bacterial genome under control of the ..., which is inducible by the addition of IPTG. T7 RNAP is then available to transcribe the gene of interest...

  21. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ...promoters for general RNAP II and RNAP III transcription; however, using viral LTRs as RNAP II promoters is ...to use an RNAP II promoter, whereas small RNAs (such as shRNA) are transcribed from the RNAP III promoters...is transcribed from DNA using an RNA polymerase (RNAP). In bacteria this is done by a single enzyme; however...of RNAs. To gain this specificity, the eukaryotic RNAP can recognize and bind to specific promoter elements...

  22. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ...A122D WPRE (AAV1) pAAV-hSynapsin1-axon-GCaMP6s-P2A-mKate2 (AAV9) pAAV-hSynapsin1-GCaMP6s-P2A-mKate2 (AAV9...signal to noise ratio and is better localized for synaptic glutamate response. They generated variants optimized...activation kinetics and localization for imaging synaptic transmission. bioRxiv 2022.02.13.480251. doi: ...AAV9) pAAV-hSynapsin1-GCaMP6s-P2A-mRuby3 (AAV9) pAAV-CAG-SomaGCaMP6f2 (AAV9) Plus, additional AAV serotypes...the most recent jGCaMP8 calcium sensors: pAAV-hSynapsin1-axon-jGCaMP8s-P2A-mRuby3 (AAV1) pGP-AAV-syn-jGCaMP8f-WPRE...

  23. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...are released in each synaptic event and they are rapidly taken up by postsynaptic receptors and transporters...sensitivity. iGluSnFR4f is well-suited for imaging rapid synaptic events such as in the somatosensory cortex, while...ideal for video recordings of large populations of synapses and deep-brain fiber photometry.  Figure...promise to be a popular choice for a variety of synaptic imaging applications. Find iGluSnFR4 plasmids ...with tools like mass spectrometry, providing a snapshot of interacting proteins at a certain time and ...

  24. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

    Type
    Blog Post
    .../B), which cleaves the synaptic protein VAMP2, a protein essential for synaptic transmission. Cleavage... Hot Plasmids podcast Light-mediated block of synaptic transmission Article contributed by Klaus Wanisch...optogenetic tool that allows light-mediated silencing of synapses was developed by Matthew J. Kennedy’s and Chandra...demonstrated the utility of light-mediated suppression of synaptic transmission in brain slices from mouse, and showed...

  25. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    ...Many thanks to our guest blogger Erik L. Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by guest blogger Erik L. Snapp. Stop using EGFP/GFP for fusion proteins! Despite...distance runner, gardner, and cook.   References 1. Snapp, E.L., et al., Formation of stacked ER cisternae... Resources on the Addgene Blog Read Professor Snapp's Post on mOX FPs Find the Best Fluorescent Protein...

  26. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...individual presynaptic terminals, which are jam packed with everything needed for synaptic transmission...it difficult to study the inner workings of the synapse by conventional light or fluorescence microscopy...researchers gain a deeper understanding of neuronal synapses, centrosomes, parasites, and more (Bertiaux, et... al (2010) Superresolution Imaging of Chemical Synapses in the Brain. Neuron 68:843–856. https://doi.org...

  27. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...fragments, expressed in the presynaptic region of one neuron and postsynaptic region of a different neuron...Currently available from the Kim lab are 2 presynaptic and 2 postsynaptic targeting mGRASP plasmids. Additionally...efforts to map the location and distribution of synapses in the mouse brain. Kim et al is utilizing a mammalian... mammalian GRASP (GFP reconstitution across synaptic partners) technique based on functional complementation...neuron, come into proximity in the synaptic cleft, functional fluorescent GFP is reconstituted in vivo. ...

  28. Tips for Getting a Faculty Position

    Type
    Blog Post
    ...  Erik Snapp just released version 3.4 of his eBook, How to Get a Faculty Position. This edition has ...along with new thoughts on research statements. Snapp, who is always looking for ways to improve his eBook...suggestions, please do not hesitate to email me at snappe@janelia.hhmi.org." You can find his original blog... Sides," are summarized below. Check Out Erik Snapp's Full "Applying for a Faculty Position" eBook Here... Skye). Good luck with your search!   Erik Lee Snapp received his PhD in Molecular Microbiology and Immunology...

  29. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ...axonal transport  (Murlidharan et al., 2014), and postsynaptic transmission (Oh et al., 2020). For example,...Others, like AAV1 and AAV9, can also transduce postsynaptic cells when used at very high titers (Zingg et...Selective promoters Selective promoters, such as synapsin, glial fibrillary associated protein (GFAP), or...Rah, J. C., & Park, H. (2020). Labeling dual presynaptic inputs using cFork anterograde tracing system...Zhang, L. I. (2017). AAV-Mediated Anterograde Transsynaptic Tagging: Mapping Corticocollicular Input-Defined...

  30. New Viral Vectors - March 2024

    Type
    Blog Post
    ... pAAV-hSynapsin1-RdLight1 AAV9 Biosensor Tian New viral vector pAAV_hSynapsin_psychLight2 AAV9 Biosensor...lab. Also of interest is the Tian lab's pAAV_hSynapsin_psychLight2 (AAV9), which contains a biosensor that...Biosensor Tian New viral vector pAAV.hSynapsin.SF-iGluSnFR.A184S AAV9 Biosensor Looger New serotype ...

  31. New and Upcoming Viral Vectors - June 2019

    Type
    Blog Post
    ...WPRE 111262  AAV1 pAAV-hSynapsin1-axon-GCaMP6s 112005  AAV9 pAAV-hSynapsin1-axon-GCaMP6s-P2A-mRuby3... Plasmid Serotype Name 106176  AAV1  pAAV.hSynapsin.SF-iGluSnFR.S72A New calcium sensors Plasmid...axon-GCaMP6s-P2A-mRuby3) 112010  AAV9 pAAV-hSynapsin1-FLEx-axon-GCaMP6s New voltage sensors Plasmid Serotype Name ...

  33. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    ...that are intricately wired into circuits through synaptic connections (Shepherd, 2004). Understanding the...something researchers often want to do at neuronal synapses for instance. Also these tags need to be fused...proteins. For example, N-cadherin (cadherin-2) is a postsynaptic cell adhesion protein that plays a critical ... PMC4573404. 2. Shepherd, G.M. (2004). In "The synaptic organization of the brain". Oxford University ...

  34. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ...Young JAT, Callaway EM (2007) Monosynaptic Restriction of Transsynaptic Tracing from Single, Genetically... can travel in a retrograde direction from the synapse to the cell body, making it a great tool to study...virus envelope, travel retrogradely, and jump the synapse to the previous neuron (Wichersham et al., 2007...

  36. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    ...to mCherry, all under the control of the human synapsin promoter. Purpose of this AAV pAAV-hSyn-DIO-hM4D...restricted to a particular cell type. The human synapsin promoter further specifies neuron-specific expression...PMC3069789. 3. Kügler S, Kilic E, Bähr M. Human synapsin 1 gene promoter confers highly neuron-specific...

  37. A Look at Addgene's QC Process

    Type
    Blog Post
    ...deposited sequence(s) in SnapGene. SnapGene annotates the plasmid maps based on SnapGene’s “Common Feature” library...

  38. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ...involved in neurogenesis, neuronal survival, and synaptic plasticity. Several studies have found reduced...stimulation to emulate optogenetic treatment of synaptic pathology." Science 347.6222 (2015): 659-664. ... Van Huijstee AN, Mansvelder HD. Glutamatergic synaptic plasticity in the mesocorticolimbic system in ...

  39. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...kinase II alpha (aCaMKII; excitatory neurons) and Synapsin-1 (pan-neuronal) promoters (Figure 1b), and sgRNAs... Neuron-specific expression is mediated by the synapsin 1 (SYN) promoter, which was superior in its ability...physiology by promoting the development of neurons and synapses as well as long-term memory formation. The Bdnf...

  40. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    ... Many thanks to our guest blogger, Erik Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at the Janelia...
Showing: 1 - 40 of 163 results