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We narrowed to 849 results for: HA

Showing: 121 - 160 of 849 results

  1. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ...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...

  2. The Importance of a Fun Workplace: Company Culture at Addgene

    Type
    Blog Post
    ... 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...

  3. Plasmids 101: Protein tags

    Type
    Blog Post
    ...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...

  4. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...EGFP into pAG Lenti CMV N-HA Puro (Addgene #236079) to create pAG Lenti CMV HA-EGFP Puro (Addgene #236081...by the Loren Looger and Kaspar Podgorski Labs now has a new version: iGluSnFR4, developed by the GENIE ...we're excited to share our very own set of empty, HA-, and EGFP-expressing lentiviral backbones designed...Choose untagged or add a versatile N- or C-terminal HA tag. Selectable and stable: A puromycin resistance...Anti-GFP [N86/38.1R] (Addgene #180084), 0.01 µg/mL Anti-HA [12CA5] - Chimeric (Addgene #198003)) and Goat anti-Mouse... loci in the genome. The modified transposon RNA has untranslated regions (U, blue) on the outside of ...12 kb — with efficiencies between 5% and 11%. It has higher efficiency and lower indel formation than ...

  5. Searchable and Sortable gRNAs for Your Next CRISPR Experiment

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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 ...

  7. Scientific Sharing in the Time of COVID-19: Databases and Resources

    Type
    Blog Post
    ... 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...

  8. Same Addgene, New Look - Why We Redesigned Our Homepage & Mascot

    Type
    Blog Post
    ...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...

  9. Seeing Red: Simple GFP Photoconversion

    Type
    Blog Post
    ... 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...

  10. Make a Splash: Notions of Scientific Impact Are Evolving

    Type
    Blog Post
    ... 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...

  11. More Data for You: Find Articles Citing Addgene Plasmids

    Type
    Blog Post
    ... 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...

  12. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    ...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...

  13. Hot Plasmids - May 2022

    Type
    Blog Post
    ...   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...

  14. Antibodies 101: Selecting the Right Antibody

    Type
    Blog Post
    ...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 ...

  15. Hot Plasmids - February 2022

    Type
    Blog Post
    ...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...

  16. Antibodies 101: Isotypes

    Type
    Blog Post
    ... 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...

  17. Addgene is Stopping Use of X (formerly Twitter)

    Type
    Blog Post
    ...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 ...

  18. Some Like it Hot: Thermostable GeoCas9

    Type
    Blog Post
    ...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 ...

  19. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    ...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...

  20. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...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...

  21. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...until 2005 [1]. Like other transposons, piggyBac™ has two components, a transposon and a transposase. The...offers a large cargo-carrying capacity (over 200 kb has been demonstrated) with no known upper limit [1]....particularly useful when the gene being expressed has threshold effects, such as with signaling molecules...transgenic cell line with the piggyBac™ system is that it has a preference for integration at regions of open chromatin...removal, which alters the endogenous DNA sequence and has been suggested to cause cytotoxicity [5]. Second,...efficiency. While the efficiency of excision with PBx has been shown to be higher than that of the wildtype...Cre recombinase, the PBx is highly specific, and has not been shown to cause genomic abnormalities when...

  22. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...mutagenesis or gene targeting Victor de Lorenzo's lab has engineered a modular mini-Tn5 vector that can be ...and the subsequent biochemical characterization, has enabled scientists to engineer a wide variety of ...green to red when it is oxidized. The Timer tool has already been used by several groups to monitor protein...imported to the mitochondrial matrix. Gottlieb’s lab has used an inducible MitoTimer reporter using a Tet-... for protein-peptide locking, Mark Howarth’s lab has developed a new tool for peptide-peptide locking....optimization through engineering, Ed Boyden’s lab has identified a variety of new channelrhodopsins for...nicknamed Chronos, is blue and green light-excitable and has faster kinetics than those of other channelrhodopsins...

  23. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...between a cell that has had one viral particle integrated into its genome and a cell that has had multiple integration...The day has arrived; you’ve painstakingly cared for your packaging cell line, prepped your DNA, transfected... be significantly higher than the same prep that has been stored in the freezer. If you plan to freeze..., you may want to consider using an aliquot that has been subjected to a freeze/thaw cycle for titering...CRISPR Genome-Wide Screening Learn How Lentivirus Has Been Used for Lineage Tracing Additional Resources...

  24. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    ...science has become increasingly restricted to academic and industrial labs, but recently there has been ...KittyBiome has been incorporated as a non-profit entity and so far most of their funding has come from ... the predictions about the effects this mutation has on locomotion in our computer model to the effects...Genetics Center (CGC) at the University of Minnesota has declared our makerspace an educational institution...

  25. Career Coaching for Scientists: Why and Where Do I Find One?

    Type
    Blog Post
    ...to define how you will know that the relationship has been successful – what are the desired outcomes? ... Dora Farkas, founder of Finish Your Thesis (who has also blogged for us) offers a free report on her ...called “Secrets to Success in Graduate School”. She has also launched an online career coaching program that...in Madison, Wisconsin she also works remotely and has experience as a scientific recruiter. Chris Humphrey...of From PhD to Life. She runs great tweet chats, has an outstanding blog and, while her PhD is in History...remotely, will travel for speaking engagements and has many areas of specialty from leadership development...

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

    Type
    Blog Post
    ...in living cells. The HA frankenbody is an HA tag detection probe. It contains an HA-binding scFv that’s...often genetically fused to other proteins, which has led to their application for basic and translational...fluorescence intensity of a tagged protein. SunTag has two components: 1) a protein of interest that’s tagged...s fused to a fluorescent protein such as GFP. The HA frankenbody works just like an antibody-based probe...of cytotoxic granules from the T cell. When a CAR has an scFv that binds cancer-related antigens, binding... Addgene blog: Read about frankenbodies, an anti-HA scFv-based probe for fluorescent imaging. Learn more...

  27. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...also the fastest test that has been developed so far.  A similar method has been optimized to detect the...Two-step-SHERLOCK has a sensitivity in the zeptomolar range while One-step-SHERLOCK has a sensitivity in...State University. Since its appearance, SARS-CoV-2 has spread to almost every part of the world manifesting...full-fledged pandemic. Containing the spread of this virus has become an utmost priority for countries around the...the outbreak ultimately flattening the curve, it has become evident that widespread testing is crucial... 83 known COVID-positive samples tested, DETECTR has a positive prediction rate of 95% and a 100% negative...

  28. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    ... dark but dissociate under blue light. This tool has now been modified by Torsten Wittmann’s lab at UCSF...genetic or pharmacological manipulations on cells, and has potential use for a wide range of applications in...monomeric, 4-5 fold brighter than mNeptune684, and has a favorable pKa of 5.6. To test out this new FP in...The increasing popularity of single base-editors has resulted in the rapid expansion of the genome editing...expansion of genome editing tools, the Reuben Harris lab has deposited a reporter plasmid that allows for rapid...

  31. CRISPR in the Clinic

    Type
    Blog Post
    ...There can be no doubt that CRISPR/Cas9 technology has been a breakthrough for the genome-editing field ...CRISPR Therapies Cancer and CAR-T therapy The FDA has already approved CAR-T immunotherapy for cancer treatment...CRISPR Therapeutics. Data shared from these trials has been outstanding, with almost all patients experiencing...desired edit, but this particular class of treatments has quite the opposite aim: to destroy. One treatment...way. Delivery, especially for systemic approaches, has its own unique set of obstacles. Thus far, several...delivery tends to be trafficked to the liver. Thus, it has been used for hATTR therapy, since TTR is primarily...

  32. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ... It was inspiring to hear how the genome project has already had a major impact on undiagnosed and rare...its work on the origin of cellular life. His lab has long hypothesized that life started with RNA and ...prokaryotic genome. Then he desribed how P. furiosus has systems to defend against both DNA and RNA viruses...technical developments. Using the Tet-on system, his lab has created a tetracycline cas9 inducible knock-out system...in a complicated conformation “not like DNA which has a boring structure.” That certainly summed up this...this conference—nucleic acid science still has many surprises for the researchers in this field. Resources...

  33. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...enormously plastic tool and has taken the scientific world by storm. While Cas9 has been most widely used to... to create specific edits in DNA, there has also been significant work on constructing Cas9 transcriptional...CRISPR activation for genes with known functions has the potential to be used in therapeutics, as it can...changes in phenotypes. For example, CRISPR activation has been used to differentiate induced pluripotent stem...creation of innovative technologies. To date his group has develop several popular CRISPR-based tools for altering...Genome-wide Screens with CRISPR/Cas9 Learn How CRISPR Has Been Used with AAV Resources on Addgene.org Find...

  34. DAP arrays for multiplex gene editing

    Type
    Blog Post
    ...Recently, Xue Sherry Gao’s lab at Rice University has developed DAP arrays for efficient multiplex base...process and release its own gRNAs from a single array, has been a top choice for multiplex gene knockout or ...proper functions to enable efficient base editing and has yet to be developed for prime editing, both and their..., which is conserved in all living organisms and has previously been used for multiplex Cas9-nuclease ...lengthy promoters, such as U6 or EF1α promoters, has limited the number of gRNAs that can be packaged ...

  35. Four Factors that Differentiate the Stem Cell Field

    Type
    Blog Post
    ...to realize that the effect of Dr. Yamanaka's work has been profound, and this is a rare instance in which...address, appeared almost incredulous that the field has exploded so quickly, citing the first clinical trial...scratch the surface of different fields. This meeting has given me an opportunity to dive a little deeper into...scientists care about patients As any grad student who has spent years studying how that one protein interacts...presenters, it was gratifying to see that Addgene has ensured that plasmid acquisition is not a rate-limiting...

  36. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    ...biology. The mistrust and anger directed at GM crops has burdened this emerging engineering technology. Monoculture... metabolism are well understood. Additional work has also identified and characterized mechanisms and ...petunias. Because of the extensive research that has come before us, it’s possible for us to combine these...plant synthetic biology. Plant molecular biology has traditionally relied on overexpression and knockouts...Nikolai Braun grew up outside of Washington D.C. and has always enjoyed exploring the outdoors and experiencing...

  37. Meet an Addgenie: Insights into Grad School, Career Paths, and More

    Type
    Blog Post
    ...considering my future path. Talking to Addgenies has helped me get a much better idea of what lies ahead...Director of Product Management Eric Perkins, who has been in many roles in Addgene since he started, said... set up for failure. Listening to Kenkel’s story has helped me realize that is not true at all. Racheal...medical school was devastating. At the same time, she has an entirely new path to discover and gives her time...finance, and while I've done that, my big takeaway has been the impact I can have on an organization beyond...was one of the biggest takeaways from my co-op. It has been so insightful to learn about different career...

  38. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...technology has revolutionized the fields of molecular biology and bioengineering, as it has facilitated...site, or protospacer, in the genome. Native Cas9 has two catalytic domains, each of which cleaves one .... When native Cas9 is complexed with a gRNA that has been truncated such that it exhibits 15 or fewer ...truncated gRNAs Importantly, the use of truncated guides has largely been demonstrated to result in decreased ...creation of innovative technologies. To date his group has develop several popular CRISPR-based tools for altering...

  39. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    ...positions. There has been a lot of engagement with the Athena SWAN Charter and it has really strengthened...their pathogens. As Patron explains it, her work has always been focused on gene transfer, from transgenes...MoClo) Kit. What does it include? Patron: Sainsbury has a mandate to share and educate and enable plant science...quantitative data for regulatory elements, which has not been available for plants. Addgene: Will the ...strengthened the network of female scientists. Twitter has been really good for that too. I’m more in touch with...

  40. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...proteins such as Green Fluorescent Protein (GFP) has likely existed in creatures such as jellyfish for...from the crystal jellyfish Aequorea victoria. It has a fluorescent emission wavelength in the green portion...protein isolated from Discosoma sp. Similar work has been done to expand the red-fluorescent protein repertoire.... Split EGFP: An alternative to FRET, split EGFP has also been used to study protein-protein interactions...A wide array of GFP-based fluorescent biosensors has been designed to detect a variety of intracellular...surface of the potential applications for GFP. It has also been used to identify particular cell populations...
Showing: 121 - 160 of 849 results