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TypeBlog PostPublishedJuly 22, 2025, 1:15 p.m.... of bioluminescence is the firefly. Along with charming nighttime displays, these insects have contributed...bright. Other luciferases have been modified to change the wavelength of light they emit or to optimize...interaction occurs, the luciferase emits its characteristic light. However, if a protein interaction brings...been used in biosensors where BRET reports on the change in conformation of a single protein. Considerations...
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Viral Vectors 101: Gamma-Retroviral Packaging Systems
TypeBlog PostPublishedAug. 6, 2024, 1:15 p.m....and γ-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G (... vice versa. Envelope plasmids may be used interchangeably. Are γ-retroviruses replication incompetent...References and resources References Swift S, Lorens J, Achacoso P, Nolan GP. Rapid production of retroviruses ...cell-based systems. Curr Protoc Immunol. 2001 May;Chapter 10:Unit 10.17C. doi: 10.1002/0471142735.im1017cs31... -
How Addgene Creates Content
TypeBlog PostPublishedNov. 17, 2022, 2:15 p.m....our mission, it’s also because Addgenies love any chance to nerd out over their favorite science topics....Addgenie expertise we can. We even have a Slack channel just for asking people if they can write or review...posts get out of date as techniques and knowledge changes and require updating or retiring. CRISPR/Cas9, ..., for example, has gone through rapid change in the past few years. Thus, we are constantly updating our... -
Antibodies 101: Buffers, Storage, and Conjugates
TypeBlog PostPublishedFeb. 1, 2022, 2:15 p.m....available in your machine before you make your purchase. Picking a single fluorophore for an experiment...overlap, or the signal "spilling over" into another channel's detection range. Though you can compensate for... University Flow Core. Fluorochromes Brightness Chart. Boston University Flow Core. Retrieved January ...flow-cytometry/files/2013/06/Fluorochromes-Brightness-Chart.pdf Syracuse Flow Core. Syracuse flow core spectral... -
Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering
TypeBlog PostPublishedMay 3, 2018, 1:48 p.m...., but they do include HEPN nuclease domains characteristic of the Cas13 superfamily. Yan et al. proceeded...Ruminococcus sp. (RspCas13d), while Konermann et al. characterized orthologs from “Anaerobic digester metagenome...Cas13a and Cas13b effectors. Like previously characterized Cas13 enzymes, CasRx also displays very high...Transcriptional Activators Learn about Cas13a (C2c2) Characterization Resources on Addgene.org: Find RNA Knockdown... -
Top Requested Lentivirus and AAV of 2016
TypeBlog PostPublishedJan. 6, 2017, 3:56 p.m....makes generation of high titer Cas9 lentiviruses challenging. This cytotoxicity also causes growth rate variability...Roth’s lab at the University of North Carolina, Chapel Hill. These can be used to investigate the effects...transgene expression across different cell types (Aschauer et al., 2013). Expression of hM4D(Gi)-mCherry ...hM4D(Gi)-mCherry in fixed tissue. References 1. Aschauer DF, Kreuz S, Rumpel S. Analysis of transduction... -
Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools
TypeBlog PostPublishedDec. 6, 2018, 1:34 p.m.... Nat Biotechnol. 2018 PubMed PMID: 30125270. Changing the way we detect viral outbreaks Article contributed...equipment. What’s more, detection is based on color changes to the assay paper, so no special equipment or ...novel peptide ligand motifs Article contributed by Michael Lemieux Listen to the SH3 domain podcast segment...biochemical processes. One of the largest and best characterized of these is the Src homology 3 (SH3) domain ... -
Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer
TypeBlog PostPublishedJuly 6, 2017, 1:59 p.m....Immunologists are coming together to tackle the challenge of curing cancer in cross-disciplinary projects...projects and collaborations. Summarized by Professor Michael Karin from UCSD: “The problem with cancer research...poster sessions During the poster session I got a chance to talk with Dr. Friedmann-Morvinski about tools...27411589. 2. Todoric, Jelena, Laura Antonucci, and Michael Karin. "Targeting inflammation in cancer prevention... -
Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection
TypeBlog PostPublishedJune 25, 2019, 12:54 p.m....overall positive charge on DNA molecules so that it can more easily cross the negatively charged cell membrane...role in bacterial evolution and is the primary mechanism by which bacteria have gained antibiotic resistance...electroporation, the application of an electrical charge to cells, to increase cell membrane permeability... -
How to Keep a Lab Notebook for Bioinformatic Analyses
TypeBlog PostPublishedJune 2, 2016, 2:30 p.m....computation work comes with new sets of challenges. One challenge with computational analyses is to keep... are doing what you are doing. Another common challenge is to keep track of what works, and what does ...that further helps you keep track of your exact changes. Alternatively, you can simply make a note in your... -
Advice for Starting a Biotech Company from a Grad Student turned Entrepreneur
TypeBlog PostPublishedJuly 10, 2014, 7:17 p.m....s Lab in 2009. From 2009 to today, Sample6 has changed focus and names, swapped leadership, and grown ...much learning, the focus of the application was changed, but Sample6 still leverages the core engineering...his co-founders raised $6 million. The biggest challenge during this time was learning to communicate effectively...better than current technology and get customers to change behavior. When you are ready to commit to the company... -
Antibodies 101: Conjugation
TypeBlog PostPublishedMarch 26, 2024, 1:15 p.m....and ELISAs. Chromogenic reactions produce a color change, so they are typically used in microscopy-based...coupling can be achieved through several different mechanisms. Most take advantage of chemical groups intrinsically...that make up the disulfide bonds between antibody chains. 3) Endogenous carbohydrates added to antibodies...References Bendall SC, Nolan GP, Roederer M, Chattopadhyay PK (2012) A deep profiler’s guide to cytometry... -
The time and cost required to make a plasmid
TypeBlog PostPublishedMarch 23, 2023, 1:15 p.m....restriction enzymes or vector purchases (Table 1). Restriction enzymes and vector purchases can raise the cost anywhere...one-off projects, or even save yourself hours by chatting quickly with the postdoc who’s really good at ...harder it is to throw in last minute experiments or changes. Fun fact: Addgene’s high-throughput approach means... -
Antibodies 101: Selecting the Right Antibody
TypeBlog PostPublishedMarch 15, 2022, 1:15 p.m.... antibody cross-reacts to, then there is a good chance that the antibody could meet your needs. Multiplex...will not only have peace of mind as you make your purchase but also someone to turn to for advice if you ...application. Fig. 1: Antibody Selection Flow Chart What does validated mean anyway? You now ...Validation! Download our antibody selection flow chart. References and Resources Additional Resources... -
Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options
TypeBlog PostPublishedMay 11, 2021, 1:15 p.m....monkeys. Surprisingly, we found that this small change made a big difference. Replacing the mCherry with...does highlight the importance of a careful characterization of transgene expression. Which tag should ...RAS-responsive adenylyl cyclase complex from Saccharomyces cerevisiae by use of an epitope addition method...Raper J, Hu X, Paré J, Bonaventura J, Richie CT, Michaelides M, Mueller SAL, Roseboom PH, Oler JA, Kalin NH... -
Visualizing Protein Turnover In Situ
TypeBlog PostPublishedJan. 16, 2018, 3:20 p.m.... studying protein turnover is radioactive pulse-chase experiments. However, these experiments are problematic...synthesis: incorporation of puromycin into peptide chains." Proceedings of the National Academy of Sciences...PubMed Central PMCID: PMC4414919. 5. Yarmolinsky, Michael B., and L. Gabriel. "Inhibition by puromycin of...Single β-actin mRNA detection in neurons reveals a mechanism for regulating its translatability." Science 343.6169... -
Celebrating 15 Years of Scientific Sharing
TypeBlog PostPublishedJan. 14, 2019, 2:08 p.m.... Joanna Brownstein (our outgoing Chair) and Dave Root (incoming Chair) and our entire supportive Board...year (12% in 2018) since 2005. We’re meeting the challenges of DNA distribution to ensure scientists everywhere...happening. The Addgene team will have some fun challenges this year and I’m excited about this, but definitely... -
Choosing the B(right)est Fluorescent Protein: Aggregation Tendency
TypeBlog PostPublishedJune 15, 2017, 2:30 p.m....protein homodimerization is characterized by an affinity of 110 µM (Zacharias et al, 2002), meaning that... proteins reported in publications are only characterized under a limited number of conditions, often ...microscopy (University of Amsterdam). He develops, characterizes and uses genetically encoded fluorescent probes... -
Giving gRNAs a Facelift - Synthetic and Beyond
TypeBlog PostPublishedOct. 20, 2022, 1:15 p.m....kits or ordered as synthesis modifications when purchasing your gRNA. Pro tip: modifications are typically...expression systems are constantly “on”. There is no mechanism to turn cutting off or on at a specific time. ...and you can opt to do-it-yourself with a kit or purchase it commercially synthesized with a label. Not ..., E., Satyapertiwi, D., Ramel, MC., et al. Photoswitchable gRNAs for Spatiotemporally Controlled CRSIPR-Cas-based... -
Summer SciComm: Preprints
TypeBlog PostPublishedAug. 16, 2022, 1:15 p.m....scientists communicate with each other: from formal mechanisms like publishing a research article, presenting...settings like lab meetings, social media, coffee chats with potential collaborators, blogging and more....read freely by anyone and are typically free of charge for authors to post. They can be assessed and commented...authors over time, providing a public record of changes, critiques or endorsements. In contrast, journal-based...