... to be activated by the DREADD actuator after it has been administered. Although we found a larger proportion...DREADD to the plasma membrane. The haemagglutinin (HA) tag is a small 9 amino acid epitope of the haemagglutinin...al., 1988). Thus, we injected AAVs carrying hM4Di-HA in the brains of mice and monkeys. Surprisingly, ... a big difference. Replacing the mCherry with the HA tag resulted in abundant expression of hM4Di in plasma... where the fluorescence is not needed, the use of HA and other short epitope tags (Kimple et al., 2013...used to reveal the hM4Di coupled to mCherry (top) or HA tag (bottom). The images show examples of dendrites... to the plasma membrane. Note that the use of the HA tag significantly improves the transport of hM4Di...

...both a fundamental driver of these principles and has emerged as a byproduct of connectivity. Between 2012...integrated with most of the tools noted above, and has a modern API. Integrations with additional tools ..., so it’s possible to re-export a protocol which has been edited back to protocols.io. Thus, RSpace becomes...Figshare. Any ELN that facilitates reproducibility has to make it easy to export data from the ELN directly... these five general purpose repositories. RSpace has been integrated with two of the five: Figshare and...additional metadata in the repository after the deposit has been made. It’s possible to associate the depositor...the repository. The ability to deposit data that has already been structured in folders or notebooks in...

... a family affair back in 2004. As Addgene has grown, it has taken effort to continue the tradition and...goofing off, the company culture here at Addgene has remained paramount. Candidates who interview at Addgene...testament to the familial aspect of Addgene. As our size has drastically increased, it would have been a tight..., even for an event where there is food and beer. Ha! We had over 60 people join the fun - including a... or binging a new Netflix series. The thing that has made all the difference here at Addgene is the emphasis...and a cooler. Chances are one of your colleagues has a frisbee at least. Use it! 8. Organize If you’re...there is something fun happening! Addgene actually has chimes outside of our break room. A ring of the chimes...

...which a good antibody has not been developed (and just because your protein has a commercially available...localization, or protease protection. Thus far Plasmids 101 has covered GFP and its related fluorescent proteins,...affinity tag to it. The FLAG, hemaglutinin antigen (HA), and c-myc tags have been the workhorses of the ...Purification Good for antibody-based purification; has inherent enterokinase cleavage site GST Large Protein...against proteolysis, but may reduce solubililty HA YPYDVPDYA or YAYDVPDYA or YDVPDYASL  1.1 Affinity... a moment), and 2 ProtA IgG-binding domains. TAP has since come to encompass several other tag combinations...necessarily want them to stick around after that purpose has been served. At this point, proteases can be your...

...Alternatively, you can select a target sequence which has already been shown to work in CRISPR experiments....experiments. Addgene’s dedication to resource sharing has led us to develop a searchable and sortable datatable ... to the fact that every gRNA listed in the table has been used effectively in a published article. This...target sequence, and model system. Even if a gRNA has been demonstrated to work in your system, it is worth...that locus. The data in the validated gRNA table has been derived from scientists’ submissions, either...

...artifacts, it has better folding and maturation kinetics than other dark fluorescent proteins, it has good signal...Technology (KAIST) and Institute for Basic Science (IBS) has developed another useful optogenetic tool that enables...facilitate future work with this goal, the Barrick lab has deposited the pDCAF3-IS5 plasmid containing a synthetic...operon originates from Pseudomonas putida CBB5 and has been tailored for expression in E. coli. This operon... contributed by Christina Mork The Murakoshi lab has developed two new FLIM-FRET acceptors, ShadowG, a... due to its very low quantum efficiency. ShadowY has a lower quantum yield and higher absorption than ...

... world has battled the COVID-19 pandemic over the past few months, the scientific community has seen a...research into this disease. A key reason this research has been able to progress so rapidly is scientific sharing...is applicable to researchers worldwide. “COVID-19 has spread incredibly fast and without any notion of ...COVID-19 Resources Canada site. To date, the portal has recruited 3,218 Canadian volunteers, and features...

...story, to get scientists excited about what Addgene has to offer, and to help scientists find what they need...homepage. Blugene, our trusted friend and mascot, has undergone a make-over as well. While in the course...Blugene needed the same thoughtful treatment. Blugene has been with us since 2011, and while you may have seen...and sharing.  One big feature upgrade? Blugene now has a full expression. You’ll mostly see a friendly smile...an experiment doesn’t go as planned. Blugene also has full arms so that they can hold and hug items closer...

... Even though GFP has been so thoroughly characterized, it turns out this protein has a few more secrets...photoconvert GFP from green to RED! This simple technique has been shown to work in plant, Drosophila and mammalian...cases, however, a GFP-tagged cell line or organism has already been made, and it would be expensive and ...is great news for microscopy enthusiasts! Addgene has empty backbones containing GFP variants used by Sattarzadeh...

... even society. One obvious way to do that is and has been to publish in journals with a high “impact factor...cited articles appearing in lower-prestige journals has risen steadily from 1995 to 2013. Considering citations...online access on scientific impact Partly, the story has to do with online accessibility. It is now about ...among the most cited. And, as scientific discourse has increasingly moved online, surely times have changed...

... how a plasmid has been used by multiple labs and see what experimental systems it has been validated ...validated in.  If a plasmid's Addgene ID # has been referenced in other publications, you'll find a link to the... and the PubMed ID # (PMID) for the article that has used this plasmid. Happy researching! If you need...

...   By Ashley Waldron Proximity-dependent labeling has become a powerful approach for proteomic mapping ...our 2019 overview). Until recently, the approach has been hampered by the need to tag each protein of ...M-CREATE) by: Brian O'Neill  The Viviana Gradinaru Lab has previously developed the multiplexed version of the...sequence knock-ins by: Lucie Wilson Le Cong’s lab has developed a method using dCas9-SSAP to knock-in long... methods for large inserts. However, this system has a much lower error rate. For a one kilobase insertion... its functions. Recently, Ravid Straussman’s lab has developed visual barcodes for clonal multiplexing...

...An antibody that has been cited thousands of times may be a safer bet than one that has never been used...work. Whenever possible, choose an antibody that has been validated in your specific application. Consider...cross-reacts with. Ideally you will find an antibody that has been validated in the species your protein of interest.... Talk with your peers. If someone in your field has used the antibody before you will not only have peace...Uhlen, 2016). Be sure that the antibody you choose has been validated using one or more of the proposed ...

...targets by: Mike Lacy Adding to their set of anti-HA frankenbodies (Zhao et al. 2019), Tim Stasevich and...iRFP713, SNAP-tag, and HaloTag fusions, while the anti-HA frankenbodies include mEGFP, mCherry, and HaloTag...Adapted from Liu et al. 2021. Anti-FLAG and anti-HA frankenbodies have a variety of applications, including...blotting, and more. Given the popularity of FLAG and HA tags, you may not even need to clone new constructs...requires a small epitope tag (FLAG is eight amino acids, HA is nine), multiple repeats of the tag can recruit...larger expression requirements (Figure 1), but Cas12a has generally been underutilized in genetic screens because...

... expressing IgM. IgM has a pentamer structure, which means each IgM molecule has a binding capacity or...function is unknown. It has a monomer structure with a valency of two and has no subclasses. IgG IgG is... appropriate isotype control. An isotype control has the same constant heavy chain domain as your primary...replaces the hinge region in IgM antibodies. IgM has no subclasses and is found mostly in blood. IgD While...

...Service since 2009. He has  developed a proprietary, low cost cloning protocol that he has used for cloning ...least 24 bp with the template and if the sequence has high GC or AT content, increase the overlapping region... most of the troubleshooting regarding this step has to be strategy specific, there are few general parameters...ratio of 1:2 (plasmid:insert), but this parameter has to be optimized according to the strategy adopted...satellite colonies and they form when all the antibiotic has been used. Don’t worry too much, just pick the big...

...communities and welcome suggestions in the comments.  X has moved significantly away from the functionality and...conversation in the scientific community. Addgene has been considering a gradual move off X as a response...conflict with our values. For this reason, our timeline has accelerated. We significantly reduced posting on ...

...Kcleave (min-1). SpCas9 has maximum activity from 35-45 °C, while GeoCas9 has maximum activity from 50...the recent discovery of GeoCas9 by the Doudna lab has opened up the field of thermophilic bacteria to CRISPR...stability in human plasma The last trick GeoCas9 has up its sleeve is its increased longevity in human...stable when incubated in reaction buffer alone, but has limited activity when incubated in diluted human ...

...You will also have to make sure that your plasmid has been incorporated into the host organism, usually..., c-Flag pcDNA3, comes from the pcDNA series. It has the CMV promoter for mammalian expression, a multiple...mammalian selection cassette (e.g. 1435 pSG5L Flag HA). You can also upgrade your basic mammalian expression...considered to be a more robust expression system and has its advantages; however, remember that you will need...the specific mutations. For these purposes Addgene has a large collection of yeast expression vectors with...

...sum of epigenetic modifications across the genome) has been implicated in disease states, and targeting ...genomic loci is possible using CRISPR, and Addgene has a number of tools for this purpose. Epigenetics began...many TALEN-based technologies, the advent of CRISPR has made targeting much easier! With the help of guide...modification at specified loci. Charles Gersbach’s lab has deposited mammalian expression constructs including...BioRender.com. Tet1 demethylase Ronggui Hu’s lab has created pdCas9-Tet1-CD for targeted cytosine demethylation...Methyltransferase 3 Alpha (DNMT3A) Vlatka Zoldoš’ lab has deposited pdCas9-DNMT3A-EGFP and pdCas9-DNMT3A-PuroR...DNA Methyltransferase MQ1 Margaret Goodell's lab has deposited pcDNA3.1-dCas9-MQ1(Q147L)-EGFP, a fusion...