...fusion protein consisting of sfGFP fused to a His-tagged ABC transporter known to bind to the HMO of interest...precisely spatio-temporally controlled as a key advantage over other inducible enzyme technologies. She ...

... Dr. Deisseroth has also generously made many of his optogenetic constructs available through the vector... at the University of North Carolina) that offer his constructs. Importantly, control viruses (not expressing...guest blogger Derek Simon! Derek P. Simon received his PhD in Cellular and Molecular Biology from the University...Mary Jeanne Kreek at the Rockefeller University.  His interests include the role of learning and memory...is also a pianist and an aspiring composer. View his blog drsimonsaysscience.org or follow him on Twitter...and for good reason. For the first time in the history of neuroscience research we have a technology that...promoter of the cre recombinase while still taking advantage of the robust expression via the promoter in the...

...affinity for calcium". Please describe any tags or fusions (e.g. His, FLAG, EGFP, etc) on your insert. Use ... have the option to indicate separate promoters, tags, or cloning information for each insert.  Encodes...insert into it. Empty backbones often contain epitope tags or fusion proteins that are appended to new inserts... indicate the gene cloned into your plasmid, any tags or fusion proteins contained in the plasmid, any...Use the drop-down menu to indicate whether the tag is on the N or C terminus and whether it was present ...

...embraces open science, self-publishing directly on his website (and often posting updates on social media... vision of passive impact,” he says, noting that his team regularly shares troubleshooting tips and works...works with other labs to help them adopt the tools his group develops. And though it took a lot of work ...  Figure 1: Fluorescence intensity of EGFP-FlavinTag in E. coli changes in response to a ~25 mT magnet...effect (many do). Fusing EGFP to the self-labeling FlavinTag made a reliable one-component magnetoresponsive... AsLOV2. After several rounds of semi-random mutagenesis and screening, the EGFP and mScarlet variants...significant gains! After a few more rounds of mutagenesis, they eventually found a version of AsLOV2 with...

...guest blogger Derek Simon! Derek P. Simon received his PhD in Cellular and Molecular Biology from the University...Mary Jeanne Kreek at the Rockefeller University.  His interests include the role of learning and memory...is also a pianist and an aspiring composer. View his blog drsimonsaysscience.org or follow him on Twitter...several groups [4, 5, 6]. These have the obvious advantage of not requiring patch cable connection to the...

...presented with a physical model of his large ribosomal subunit, he kept sliding his palm across the surface of... about Jacob’s initial reaction to this model on his blog. Or, for a description of how effective this... of proteins using 3D printing technology. The tagline for our company is “…where molecules become real...Addgene.org Visit Our CRISPR Guide Page CRISPR/Cas9 History Browse All CRISPR Plasmids and Resources ...

... Cloning of basic parts (promoters, UTRs, genes, tags, etc.) into MoClo-compatible vectors. Level 1: Assembly...this cloning technique! Many thanks to Rob Hurt for his help on this piece. Rob is postdoc in Peter Schultz...

...guest blogger, Erik Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology and Immunology from...from Oregon Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz...Research Campus of the Howard Hughes Medical Institute. His interests include the quality control of secretory...Growth of new livers in culture could resolve this shortage." What is the knowledge gap you plan to address...

...epitope-targeting antibody that is conjugated to a biotin tag but not a fluorophore. In a separate step, cells ...interest. Indirect labeling is achieved through protein-tag or protein-protein interactions, like biotin-streptavidin... lab of Arlene Sharpe at Harvard Medical School. His projects include investigating negative regulators...target marker, allowing for its detection. The advantage of this system is that streptavidin conjugates...the primary antibody (Figure 1b). As above, the advantage of this system is increased flexibility for panel...of such targets include transcription factors, histones, and DNA repair enzymes. Note that when using ...stepwise decrease in fluorescence intensity in a (histogram) flow plot (Figure 4b). Through this method, differences...

...promoter. The cloning procedure adds both V5 and His6 epitope tags to the C-termini of Met proteins, which can...) from the two yeast species into the pYES2.1/V5-His yeast overexpression vector. This plasmid contains...

...out yet?) your research. Derek P. Simon received his PhD in Cellular and Molecular Biology from the University...Mary Jeanne Kreek at the Rockefeller University.  His interests include the role of learning and memory...is also a pianist and an aspiring composer. View his blog drsimonsaysscience.org or follow him on Twitter...reinforcement are ICSS and place preference.   Historically, ICSS was done by implanting an electrode and...technology is improving and may one day reach the sophistication of mouse transgenics. Everything that has been...the ferrule implants and patch cables. One huge advantage of using rats is their superiority as behavioral...

...jump into industry.” In his article On Leaving Academe, Terran Lane is asked by his colleagues and friends...learn about the diverse options and how to take advantage of them early is one way to smooth your path to...

...University studying mammalian immunology, and in his free time, he is pursuing a plant engineering project... cell culture and transgenesis he wants to share his knowledge with the community by teaching a DIY plant... entrepreneurs using our space to perform early-stage experiments. For example, Dave Grawoig is using ...

...tRNA ( 68307 ) at the central position. Introducing Hi-P for screening phosphorylation-dependent protein-protein...68295 GFP E17TAG Plasmid 68296 GFP S2TAG Plasmid 68297 GFP Q157TAG Plasmid 68298 GFP S2TAG/E17TAG Plasmid...Plasmid 68299 GFP E17TAG/Q157TAG Plasmid 53225 MBP-MEK1 S218TAG/S222TAG ( aka. PCRT7 tetR pLtetO MBP-MEK1 XX...68301 MBP-MEK1 S218TAG Plasmid 68302 MBP-MEK1 S222TAG Plasmid 68305 Beta lactamase S68TAG Plasmid 69118...-EFSep) Plasmid 34624 SepOTSα tRNA-Sep ( aka. pCAT112TAG-SepT) Plasmid 68283 SepOTSβ Plasmid 68284 SepOTSγ...69118 E17TAG GFP zeo resistance Strain 34929 BL21ΔserB Strain 52055 EcAR7 Plasmid 112031 pNAS1b split mCherry...

... for the CZ Biohub project by Manuel Leonetti to tag all 22,000 protein coding genes in the human genome... at the Biohub in San Francisco. Before starting his career in software, he completed a MSc in Psychology... tools which were optimized for HDR, except for TagIn. HDR requires the additional step of mutating guide...cut-to-insert distance for HDR. (The notable exception is TagIn––see their "summarisation score".) For CrispyCrunch...HDR. Optimal mutation in-depth One the most sophisticated features of CrispyCrunch is how it mutates guide...

...-28b-Cas9-His 6xHis (C terminal); NLS (C terminal) 62933 pET-Cas9-NLS-6xHis 6xHis (C terminal...Plasmid ID Plasmid Name Tags 69090 pMJ915 MBP (N terminal); 6xHis (N terminal); 2xNLS (C terminal... be selected, since it cleaves at the stop codon TAG. Design your repair template with 55-100 base...; NLS (C terminal) 62934 pET-NLS-Cas9-6xHis 6xHis (C terminal); NLS (N terminal)   Easi-CRISPR...

...’re attending SfN2019, stop by booth #2163 to say hi and learn more about Addgene’s viral service. Additional...topic as shown in the graphic below. Follow the hashtag #AddgeneGoesViral to find information on the basics...

...expression of the HIS3 gene, which is required for histidine manufacture and therefore growth on his-deficient...Post About Yeast Vectors Read our Blog Post About Tagging Yeast Genes ...activator (1a) or a repressor (1b). Scientists took advantage of these discoveries to create a new system for...that repress instead of activate transcription. The HIS3 reporter system (e.g. the Gateway-compatible bait... is an example of the former. In this system, histidine-auxotrophic host strains are used to select for...subset based on a cellular compartment or growth stage). A prey library can either be applied to the bait...

...promoters, terminators, solubility and purification tags, connectors, cDNAs, and more to find the perfect...constitutive and inducible promoters, linkers, and tags makes EcoFlex a great option for those interested... a larger library of parts but don’t need to add tags to your proteins, then the CIDAR Kit is an excellent...Stroik and Meghan Rego, with thanks to Rob Hurt for his expertise and input.  References and resources References...

...Neuroscience Conference 2023? So are we! Come say hi to us at Addgene's booth, #3422, in the nonprofits...also be sharing our conference experience on X, Instagram, and Bluesky.   Finally, if you're looking to ...

...resources. Listen to hear all about Professor Goedhart, his lab, and some of the tools he’s developed.   Listen...creation of improved fluorescent proteins through mutagenesis. Chapter 6 - The future of FRET and fluorescent...

...guest blogger, Erik Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology and Immunology from...from Oregon Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz...Research Campus of the Howard Hughes Medical Institute. His interests include the quality control of secretory... be needed to achieve your intended next career stage and create a timeline for achieving these goals....publications (published, not in preparation) A history of successful funding (fellowship grants) A project...

... in the making by Dr. Zhuhao Wu and his colleagues! Dr. Wu and his lab members (like me!) are dedicated...compatible with the desired applications (such as immunohistochemistry (IHC) or western blot), and cost-effective...rAbs are a relatively new technology that takes advantage of DNA sequencing to turn mAb clones into expression...

...supervisor’s management style, bring challenges to his or her attention as soon as you can. It is in your...you communicate your ideas assertively, listen to his or her viewpoint, and discuss options for mutually...people: Hostile-aggressives, who are usually antagonistic and impolite, Complainers, who spend most ...

...Cruz where his group focuses on gene regulation in nematode molting and spermatogenesis. His group also...conditionally manipulate proteins. Learn more at his webpage or follow him on Twitter.   References Arribere...-CRISPR and co-conversion approaches have the advantage of being used in any genetic background, but require...

...measurable through an epitope tag such as FLAG/His/GFP. Fluorescent protein tags, which correlate closely ...different epitopes. Detection of overexpressed (tagged or untagged) protein When expression plasmids are available...population (pink histogram) compared to the WT cells (green histogram). The unfilled histograms show cells ...400,000 cells were stained with the indicated Synaptotagmin-1 antibodies and corresponding Multi-rAb CoraLite...You can see an example of their work on their synaptotagmin 1 page. The data produced is available on the...interpretation of your results. Therefore, taking advantage of antibody validation efforts and information...to selecting high-performing antibodies for Synaptotagmin-1 (Uniprot ID P21579) for use in western blot...

...Matthijs Dorst’s Deck the Lab Nativity scene below his Christmas tree! It is that time of the year again...showcase it all!           View this post on Instagram                       A post shared...dyed with real microbial pigment. Check out the hashtag to see the description of all the ornaments. #DeckTheLab...

...Mammalian Sato 123128 pET-His-Cre 6xHis-Cre T7 Bacterial Liu 123131 pET-His-VCre 6xHis-VCre T7 Bacterial Liu...hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR Cre, KASH-tagged EGFP, and sgRNA expression hSyn AAV Zhang 60820...9xGGS-Cre-6xHis Cre fused to highly cationic GFP variant T7 Bacterial Liu 62939 pET-Cre-6xHis Cre-6xHis T7 Bacterial...73472 RabV CVS-N2c(deltaG)-mCherry-P2A-Cre Cre CMV RABV Jessell 73474 RabV CVS-N2c(deltaG)-Cre Cre CMV RABV...Simpson 106368 pCMV-Tag2B-NCre N-terminal split-Cre CMV Mammalian Hirrlinger 106369 pCMV-Tag3B-CCre C-terminal...106370 pCMV-Tag2B-NCreERT2 N-terminal split-Cre-ERT2 CMV Mammalian Hirrlinger 106371 pCMV-Tag3B-ERT2CCre ...homology arms Mammalian Heller 113837 pCAGGS-mTagBFP2-T2A-iCre TagBFP, iCre CAG Mammalian Capecchi 113849 pCAGGS-pac-T2A-iCre...

...innocuous tags” (inntags) less likely to alter a protein’s properties. Why do we need new epitope tags? Short... looking to a tag a protein, inntags IT5 and IT6 are a great place to start. These tags are easy to use... about other Protein Tags in our Plasmids 101 Series. Get the Inntag Plasmids Tag endogenous proteins ...major caveat - a tag may interfere with protein localization and/or function, so each tagged protein must ...proteins found that 20% of tagged proteins did not localize similarly to their untagged counterparts. These ...the effects of GFP fusions to inntags or traditional FLAG, HA, and Myc tags. Impressively, IT6-GFP did not...of the traditional tags were suitable for both applications. To prove that inntags are indeed innocuous...

..., modular plasmids, combining fluorescent tags, epitope tags, protease sites, and selection markers. These...fluorophore tagging vectors for your single or multi-color imaging experiments. Interested in epitope tags? Others...have genome tagging systems enabled you to advance your research? Do you have a favorite tagging system that...epitope tag (12xHis, 2xStrep, 3xFlag, Protein A, or V5), and selection marker (KanMX, HphMX, or His3MX).  ...Researchers have long taken advantage of this natural process to integrate protein tags into the genomes of S...These tagging vectors are based on previously described pFA6a-link vectors and include a Kan, SpHIS5, or...especially useful in protein complex studies where tagging of multiple protein products is desired, as multiple...