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Showing: 481 - 520 of 879 results

  1. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 3, 2022, 1:15 p.m.
    ...popular split-FP plasmids from our catalog to help you find the right split-FP plasmid for your experiment.  ...Browse the table below, and click the Article link to find all the plasmids from the associated paper:   ... a protein fusion may disrupt functionality or binding of your target proteins, especially if the termini... principles and experimental design. We hope you find these plasmids useful. Good luck, and happy experimenting...peroxidase (Alice Ting Lab 2016)  Split-Cas9 for inducible genome editing (Feng Zhang Lab 2015)  Reference...

  2. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

    Type
    Blog Post
    Published
    May 4, 2021, 1:15 p.m.
    ... plasmid. (Right) An assembled antibody protein binds an antigen. Image from Fvasconcellos.     How...Each antibody would then be tested for antigen binding. If starting from an established hybridoma line...and maturation in vitro by selecting for antigen binding from libraries of recombinant antibodies. The selection... and producing antibodies from plasmids is much kinder to our animal friends! Finally, if we want better... Leonhardt H (2015) Nanobodies and recombinant binders in cell biology. Journal of Cell Biology 209:633...

  3. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...takes about one day, even for six plates (576 individual plasmids)! These libraries are then QC’d, pooled...the raw data from each sample in our pool and individually assembles the reads into a single FASTA sequence...necessarily expecting a perfect match - we will often find a few mismatches in the origin of replication or...insertions that could compromise function. When we do find mutations, we check to see if they affect the translated... detecting common features using Snapgene. If we find information that differs from what we would expect...

  4. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    Published
    Aug. 6, 2019, 12:55 p.m.
    ... used to separately introduce each mutation independently into different mice, and then these strains ...generations as well, a concept known as linkage. This hinders the likelihood that mice in the F2 generation and...may also be either constitutively expressed, or inducible (this is explained more in Part 1 of this series...PMC4448610. Additional resources on the Addgene blog Find our Plasmids 101 blog posts Read blog posts about...engineering blog posts Resources on Addgene.org Find more about Cre-lox Read our Genome Engineering Guide...

  5. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    Published
    Aug. 1, 2024, 1:15 p.m.
    ...stronger binding antibodies will require harsher stripping to remove, so start with the weaker binding ones...stripping solution uses low pH to disrupt antibody binding to antigens. Ingredient Concentration or... time will depend on your blot — antibodies that bind more strongly or that have saturated the blot may... the Addgene blog The Basics of Western Blotting Find other tips and tricks for molecular biology protocols...

  6. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    Published
    Aug. 7, 2018, 12:31 p.m.
    ...expression in your target cell line, researchers often find themselves faced with a roadblock. In this post,...bit more challenging. With these considerations in mind, when planning an experiment users should weigh ...the best for others. For example, in our hands we find that a double harvest approach at 48 and 72h produces...collected from an unconcentrated prep. With this in mind, if the experiment requires a certain volume of ...page Resources on Addgene.org Lentiviral Guide Find lentivirus plasmids Browse all viral vectors ...

  7. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    Published
    July 11, 2023, 1:15 p.m.
    ...chemogenetics in our chemogenetics guide Find viral preps for your research Find chemogenetics plasmids for your...you choose, there is no “perfect” drug that only binds to one receptor, so it is important to fully characterize... ligands. PSAMs themselves are modified ligand binding domains. When coupled to the ion pore domain (IPD...Ligand-Dependent Analgesia, as well as Ligand-Independent Functional Changes in Sensory Neurons. The Journal...

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

    Type
    Blog Post
    Published
    Dec. 1, 2015, 3:30 p.m.
    ...over 300 Chinese scientists eager to hear current findings in nucleic acid science. I was delighted to see...if the bags contained flora or fauna. I tried to find out what certain ingredients might be used for (...Molecular Medicine presented another unexpected finding in the regulatory RNA field. The Rajewsky lab is...system, his lab has created a tetracycline cas9 inducible knock-out system. His lab is also exploring the...Thomas Steitz of Yale. He gave an update on new findings from structural studies on the ribosome performed...

  9. Gaining Leadership Skills Volunteering at a Professional Organization

    Type
    Blog Post
    Published
    Aug. 6, 2015, 2:30 p.m.
    ...develop new working relationships with these individuals who often had different styles from their predecessors...laid out plan of action, others would act more independently and some others would need regular re-direction...organization speaks in “one voice” whilst among individual members, many opinions may exist. I learnt to...Becoming involved is not too difficult and you can find opportunities around you. You just need to be prepared...Twitter @JulietAdvocSci.        Further Reading Find more Career articles on the Addgene Blog Read Joanne...

  10. Transferable Skills: Negotiation

    Type
    Blog Post
    Published
    March 3, 2022, 2 p.m.
    ... suits discussing multimillion dollar contracts behind closed doors. Fig. 1: The mythical land ...get what I want, that means you don’t. In this mindset, one of us wins, the other loses. Although this...examples depend on many specific details about the individuals, each example provides a scenario where both ...to you being remembered as a person who tries to find solutions that benefit everyone. Even if I “lose...positively if they tried to understand my position and to find a creative win-win solution. What does professional...

  11. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ... I’m not going to lie, AAV transfer plasmids are kind of a pain to work with. These plasmids contain ITRs...digests. I freeze the bacterial pellet for the remainder of the cultures and later maxiprep only the cultures...18265393. Additional resources on the Addgene blog Find out what makes AAV a versatile tool for gene expression...calculator for planning AAV packaging transfections Find tips for a first time CRISPR user Resources on ...and lentivirus Check out Addgene's virus protocols Find AAV for your research ...

  12. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    Published
    Oct. 11, 2018, 12:30 p.m.
    ...techniques Resources on Addgene.org Find basic molecular biology protocols Find plasmid cloning protocols Check...upon growth on LB/Cam/X-gal/IPTG agar plates, indicating successful reconstruction of the coding sequence...assembly that would result in a blue phenotype. Indeed, this was seen in all lacI/ lacZ assembly test ... efficiencies and accurate assembly levels were indeed possible, with correct, in-frame assembly proceeding...

  13. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    Published
    Sept. 6, 2016, 2:30 p.m.
    ..., Ras proteins have stumped the pharmaceutical industry for years. They have proven very difficult to ... lack areas that are accessible, high-affinity binding targets for potential therapeutic agents. Nevertheless...demonstrated that a number of novel small molecules can bind directly, albeit weakly, to K-Ras and inhibit its... in human cancers, and synthesizing compounds to bind specifically to their mutated residues. For Ras ...Sun, Qi, et al. "Discovery of small molecules that bind to K‐Ras and inhibit Sos‐mediated activation." Angewandte...

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

    Type
    Blog Post
    Published
    April 19, 2016, 2:30 p.m.
    ...Fluorescent Protein Resources Find Other Plasmids that Glow (Luciferase Plasmids) Find Optogenetics Resources...protein. A dimeric fusion protein will have two FP binding domains forming a building block sufficient to ...your studies and greatly lower your chances of finding artifactual results. Many thanks to our guest ...Addgene Blog Read Professor Snapp's Post on mOX FPs Find the Best Fluorescent Protein for Your Needs Get ...

  15. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    Published
    May 20, 2014, 2:06 p.m.
    ...available FP emission colors and 10 points to keep in mind when choosing an FP (or two) for your upcoming experiments... used are GECIs - genetically encoded calcium indicators (e.g. GCaMP). Others include: pHluorin & pHTomato...Fluorescence Complementation (BiFC). 8 points to keep in mind when choosing a fluorescent protein Excitation ...considered when labeling proteins that interact. Indeed, FRET is often used to determine if two proteins...isolated from the Unagi eel was shown to mature independently of oxygen, making suitble for use in anaerobic...

  16. Great Results Start with Great Standard Curves

    Type
    Blog Post
    Published
    Aug. 15, 2024, 1:15 p.m.
    ...rely on standard curves in their day-to-day work. Indeed, viral vector production facilities frequently ...diluted in a series ranging from 1:2 to 1:32. Each individual step of the series is consistent at 1:2. When... from 0 to 1 with 1 being a perfect fit and 0 indicating that there is no linear relationship between ...the %CV of the standard curve, one averages the individual %CVs for each point on the curve. ‌The smaller...complex trendlines. If you don’t have access to this kind of software, don’t give up! Rather than plotting...

  17. Getting to Know Expansion Microscopy

    Type
    Blog Post
    Published
    June 20, 2023, 1:15 p.m.
    ...the inspiration behind ExM, how it works, and some of the limitations to keep in mind whether you are ... within ~250 nm of each other. For reference, individual presynaptic terminals, which are jam packed with...the equipment needed for EM is not something you find in the average cell and molecular biology lab, and...their relative positions, allowing you to resolve individual points with a conventional, diffraction-limited...

  18. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    Published
    Dec. 22, 2022, 2:15 p.m.
    ...donor get used as a template for repair? Read on to find out! Homology-directed repair and genome engineering... to form ssDNA and uses the ends of the ssDNA to find a repair template. The highest HDR efficiency is...clones by reading up on validating genome edits to find screening strategies for your experiment. Also learn...homology-directed repair choice in response to CRISPR/Cas9 Induced-double strand breaks. Int J Mol Sci., 21(18): 6461...Resources on Addgene.org Browse All CRISPR Plasmids Find Validated gRNAs for Your Next Experiment Check out...

  19. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    Published
    May 19, 2016, 2:30 p.m.
    ...Andre K, Stein D, Blick G, Greenberg RN, Kinder C, Zolopa A, Binder-Scholl G, Tebas P, June CH, Humeau LM...addition of a chimeric 5’ LTR containing a Tat-independent promoter. Although safer, this system may be ...839-48. PubMed PMID: 26758916. 7. Transfusion independence and HMGA2 activation after gene therapy of human...Our Lentivirus Guide Learn about Virus Biosafety Find Lentiviral Vectors for Your Research  ...

  20. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ...Therefore, the roles/functions of individual proteins can be studied independently of other viral proteins simplifying...this would prevent the study of the proteins independently from each other (see Uses of Minigenome System...luciferase can readily be measured to quantify (indirectly) transcription performed by the viral polymerase...easily studied as each component can be examined independently. In addition, experiments can be performed in...

  21. Plasmids 101: Protein tags

    Type
    Blog Post
    Published
    Dec. 11, 2014, 4:26 p.m.
    ...calmodulin binding peptide (CBP), a TEV cleavage site (more on that in a moment), and 2 ProtA IgG-binding domains...with folding and stabilization, as can maltose-binding protein (MBP). Overexpression can also lead to ...KRRWKKNFIAVSAANRFKKISSSGAL 4 Affinity and Purification Binding and elution steps use very moderate buffer conditions...proteases without requiring the addition of an independent cleavage recognition site. In fact, SUMO cannot...

  22. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    Published
    Sept. 23, 2014, 3:05 p.m.
    ...month it’s been viewed by users in the US, China, India, Canada, the UK, South Korea, and beyond!   Check...use, and add your own comments. Currently you can find protocols from NEB, OpenWetWare (see below), and...classic protocols on Protocol Online that you might find useful for your research. LabGuru Website: http:...pricing options for academic labs, departments, industry, and more to fit every lab's needs. Plus there...offers software designed with scientists workflow in mind and can offer a one-stop solution to your lab's ...

  23. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    Published
    Nov. 12, 2020, 2:15 p.m.
    ...synthesis relies on transfer RNAs (tRNAs). tRNAs bind to and deliver amino acids to the ribosome where...complementary to the mRNA sequence. The 3’ end of  the tRNA binds the amino acid that corresponds to the anticodon...1980s, scientists even created a codon adaptation index (CAI) that is based on the codon usage frequency...https://doi.org/10.1038/nsmb1031 Gustafsson C, Govindarajan S, Minshull J (2004) Codon bias and heterologous...nrg3051 Sharp PM, Li W-H (1987) The codon adaptation index-a measure of directional synonymous codon usage ...

  24. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    Published
    April 20, 2021, 2:30 p.m.
    ...different levels of cytotoxicity and the mechanisms behind the cytotoxicity can vary. For example, the popular...virus G protein. VSV-G is a trimeric protein that binds phosphatidylserine and low-density lipoprotein receptors...the potential for off-target infectivity as it can bind and transduce many cell types before reaching the...needs. Check out some of these review articles to find tables and information on different viral membranes... on Addgene.org: Check out the lentivirus guide Find ready-to-use lentiviral preps at Addgene ...

  25. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ... everyone. Addgene is certainly going to have to find ways to tag and curate reagents from these emerging...emerging organisms so scientists can easily find the useful new reagents that come out of this work as the...parenting strategies and make important ecological indicators since species live in water and on land. Current...nests so can be studied over time. They recognize individual faces (I guess of other wasps) so can be used... expensive. They are using a cool fluorescent indicator system to rapidly identify recombinants. Arthropods...

  26. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    Published
    March 12, 2019, 1:37 p.m.
    ...committee to understand your goal, what you expect to find, and how you will interpret and use the information... correct or your data favor a different model. Finding evidence for or against a model is a standard scientific...knowledge gap? This includes the overview and the individual aims. Describe the question, the bare minimum...know they will work. Stabilize your blood sugar. Find an energy bar that you like and consider eating ...photographer.  Additional resources on the Addgene blog Find tips for presenting a science talk Check out our...

  27. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    Published
    July 30, 2024, 1:15 p.m.
    ...Addgene’s 20th anniversary celebration, we’ve been finding and sharing stories unique to our repository and...biosensors were distributed in 2023 alone. You’ll also find a large number of requests for vectors in the “controls...available in the repository. GCaMP, a calcium indicator (biosensor, by Addgene’s categorization) was first...community they serve. As difficult as it may be to find answers to such questions, Addgene is deeply interested...will continue to do so, in the hopes that these findings can help others as much as they've helped us.  ...

  28. How to Write a Scientific Review Article

    Type
    Blog Post
    Published
    Feb. 16, 2017, 3:30 p.m.
    ...I mentioned, I wasn’t an expert in the field. To find reviews, I just searched online and found ones that...well-known researchers in the field. I made sure to find information from genome-wide studies, as well as...you need to read every paper from start to finish. Find the information that’s relevant to you, note it,...could see how big the studies were, and you could find the original paper if you wanted to learn more. ...look done, just keep working on it. Also bear in mind that this is just a review article and not your ...

  29. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    Published
    Nov. 3, 2016, 2:30 p.m.
    ... to everyday obstacles for many of us, such as finding time to read all of these articles. Most of us ...research. So how can we fault others for failing to find our work?  I study processes for diagnosing and ...where you can see the yearly interdisciplinarity index of research for single or multiple disciplines –... more, you can then link to the journal article, find blogs about the study, or even review Facebook and...aquaculture/ The goal of the WeShareScience project is to find new and exciting ways to share ideas derived from...

  30. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    Published
    Oct. 25, 2016, 2:30 p.m.
    ...information for postdoctoral positions is very hard to find and this is partly due to the administration of ...paid. Institutional salary policies are one indicatator of how salaries may changes due to the FLSA. ...Current salary information that we have been able to find suggests that salaries may even double. The University...to the new level:  We are still waiting for an indication of what (most of) Harvard, Johns Hopkins, (most...make gradual adjustments towards this new level. Indeed some institutions did this. It doubtless took a...

  31. The Challenges of Cell Culture

    Type
    Blog Post
    Published
    May 5, 2016, 2:30 p.m.
    ...Proliferation and Invasion in Renal Cancer Cells, Marker-independent identification of glioma-initiating cells. The...that can impede collaborations, especially with industry. Today we also have the appropriate tools to verify... standards and has laboratories to carry our independent quality control for a range of testing, from ...forensics, DNA sequencing to drug testing. You can find Nick on LinkedIn. References 1. Skloot, Rebecca,... Protein Localization Resources on Addgene.org Find Plasmids for Mammalian Cells ...

  32. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    Published
    April 7, 2016, 2:30 p.m.
    ...other important binding events take place during the course of infection. These binding events can be impaired...infect and select, seems simple, in actuality, many find using lentivirus to be time consuming, difficult...possible. One problem that often plagues labs and hinders experiments is mycoplasma contamination. Mycoplasma...Lentiviral Vectors Additional Resources on Addgene.org Find Viral Vectors for Your Research Brush Up on All ...

  33. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    Published
    Oct. 1, 2015, 2:30 p.m.
    ...nose-poke at a particular hole (the active hole) induces laser stimulation of the chosen brain circuit, .... If activation of the circuit is sufficient to induce reinforcement (instills a sense of “reward”), then...software such as EthoVision. Consult the literature to find all the applications of optogenetics relevant to...region may affect the animal; stimulation could induce seizures or some other unwanted phenomenon or it...learn optogenetics is straight through the source - find a lab within your own neuroscience sub-discipline...

  34. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    Published
    Nov. 5, 2014, 3:41 p.m.
    ...hero (that I’m not), FRET loves playing “live.” Indeed, FRET was one of the first techniques which enabled...Historically, molecular interactions were detected by indirect means often using probes with the potential to...may have to design several different constructs to find one that could be used for the purpose of your studies.... But it is a parameter that you have to keep in mind when analysing FRET results. The cross-talk between...also don’t have any resonance transfer. You have to find a balance between FRET efficiency and cross-talk...

  35. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    Published
    March 27, 2018, 1:32 p.m.
    ...University of Washington reported a surprising finding - when delivered via AAV infection, gene targeting...efficiency gene targeting approaches improved on this finding with the addition of selectable markers that are... et al 2004, Kim et al 2008). While the biology behind this AAV effect remains unclear, it is likely not...infect recipient cells. After G418 selection, individual gene edited clones are identified by PCR. Then...Resources on Addgene.org Check out our Viral Service Find AAV Vectors Browse all Viral Vectors ...

  36. 10 Steps to a Perfect Science Talk

    Type
    Blog Post
    Published
    Aug. 23, 2016, 2:30 p.m.
    ... Joanne's 10 Things Scientists Ask about Finding an Industry Job Talk What’s Your Story? By Herminia Ibarra...speaking early and often. Two of my jobs have indirectly resulted from someone seeing me speak in a non-interview...before tailoring each presentation appropriately.  Find out who will be listening.  Are they early, mid,...explanation. The people that need the information will find it. Experienced speakers present useful protocols...

  37. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    Published
    Nov. 7, 2014, 2:56 p.m.
    ... of E. coli used today are descended from two individual isolates, the K-12 strain and the B strain. K...used in the lab Most of the commercial strains you find today are marketed for a specific purpose: fast ...conjugation. Genes listed on F´ are wild-type unless indicated otherwise fhuA (formerly tonA) ferric hydroxamate...typically standard; just having the gene listed indicates it is non-functional. If a gene is deleted, it...Vectors Looking for More Plasmids 101 Articles?  Find them all at our Plasmids 101 Topic Page!     ...

  38. Introducing the Addgene Antibodies 101 eBook!

    Type
    Blog Post
    Published
    Oct. 29, 2024, 1:15 p.m.
    ...’t quite seem to find an antibody that works, we highly recommend “Chapter 3: Finding the Right Antibody...very new to antibodies as research tools, you may find “Chapter 1: What is an Antibody” quite useful. If...place to start looking for answers.    We hope you find the latest addition to our eBook collection useful...

  39. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    Published
    Jan. 16, 2025, 3:42 p.m.
    ...for your experiment? Use Addgene’s AAV Data Hub to find experimental reports on commonly used vector cargo...Serotype 4 (AAV4) and AAV5 Both Require Sialic Acid Binding for Hemagglutination and Efficient Transduction...Viswanathan, S., Gaj, T., Lavzin, M., Ritola, K. D., Lindo, S., Michael, S., Kuleshova, E., Ojala, D., Huang..., M. J., Chiorini, J. A., & Zabner, J. (2001). Binding of adeno-associated virus type 5 to 2,3-linked ...α2,6 N-Linked Sialic Acids Facilitate Efficient Binding and Transduction by Adeno-Associated Virus Types...

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

    Type
    Blog Post
    Published
    April 5, 2018, 12:42 p.m.
    ...Addgene.org Check out our CRISPR Guide Find Plasmids for Cas9 Purification Find CRISPR Plasmids for Mammalian ...including one loxP site insertion and one NHEJ-repaired indel, often occur. Figure 1: In conventional gene...recombinases (Cre, Flp etc.), and tetracycline drug induction systems (ttA or rtTA) are between 1 to 2 kb in...al. saw success with Easi-CRISPR across three independent core facilities for 13 projects, with most projects...
Showing: 481 - 520 of 879 results