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TypeBlog PostPublishedJan. 24, 2019, 2:24 p.m....stochastic and combinatorial expression of fluorescent proteins is only possible at low copy number ranges, resulting...2018) (Figure 1). Once introduced, fluorescent proteins were best imaged after tissue clearing with SeeDB2...expression levels. Clearing large tissues - Fluorescent proteins are very stable in SeeDB2. However, SeeDB2 is ...strategies for combinatorial expression of fluorescent proteins in the nervous system." Nature 450.7166 (2007)...labeling of neuronal circuits with fluorescent proteins and chemical tags." eLife 7 (2018): e40350. PubMed...
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What the HEK?
TypeBlog PostPublishedApril 18, 2023, 1:15 p.m....recombinant proteins at high levels. In a case where clinical applications of recombinant proteins are necessary...cells may be particularly useful due to their intrinsic properties. HEK-293 cells are tumorigenic, ...cell line’s ability to faithfully produce these proteins and its relatively low expression of native channels...Cell Line as a Platform to Produce Recombinant Proteins and Viral Vectors. Front Bioeng Biotechnol. (2021... -
Hot Plasmids and Viral Preps - September 2021
TypeBlog PostPublishedSept. 21, 2021, 1:15 p.m....-free system, capable of producing fluorescent proteins and enzymes. Image from Moore et al., 2021. ...through monitoring the expression of fluorescent proteins and oxytetracycline enzymes. Next, this system...libraries: Cas13X and Cas13Y, two compact Cas13 proteins found in the metagenome datasets from hypersaline...messenger-RBP sgRNA Library targets human mRNA binding proteins with 10 guide RNAs per gene. The lab modified ...also target core operons in other pneumococcal strains. The BARBEKO sgRNA library uses CRISPR cytosine... -
Donations from Addgene to Yield Answers for Rare Disease Researchers
TypeBlog PostPublishedSept. 15, 2015, 2:30 p.m....Every year, the Rare Genomics Institute sponsors a global competition for researchers working on rare...He says that the award from the Rare Genomics Institute and Addgene will make a substantial difference... (top right) and his colleagues at the Wistar Institute in Philadelphia, PA will put their award to work...Joshua Wang (bottom right), also of the Wistar Institute, says that the lab got interested in XP out of...Resources Find out more about the Rare Genomics Institute and the BeHeard Award! ... -
What Good Citizenship Can Do for Reproducibility in Science
TypeBlog PostPublishedNov. 7, 2019, 2:18 p.m.... when a researcher decides on a new project and begins by trying to reproduce someone else’s result. Then...time that they thought would be moving forward instead trying just to get going. It’s like being stuck... in science needs both a job and funding, so institutions and funders, to a large extent, hold the keys...is clearly room to do more. If we could really instill a culture of rewarding good scientific citizenship...UK and then a postdoc at The Scripps Research Institute in La Jolla, California. She joined Elsevier after... -
Reagent Repositories Are Speeding up Science During the Pandemic
TypeBlog PostPublishedAug. 4, 2020, 1:15 p.m.... Biological repositories that distribute mouse strains, plasmids, cell lines, and much more have been ... Bioresources Cell Bank, part of the National Institutes of Biomedical Innovation, Health, and Nutrition...19 plasmids and resources! Case study 3: Mouse strains Reagents developed during the SARS-CoV pandemic...The Jackson Laboratory, a nonprofit research institution, has been able to rapidly propagate this strain... Aliyah Weinstein... -
Quantifying DNA? Here are Five DNA Quantification Methods to Consider
TypeBlog PostPublishedJuly 21, 2020, 1:15 p.m....based on your downstream application, time, and instrument availability. UV absorbance Using UV absorbance...acids (DNA and RNA) absorb maximally at 260 nm. Proteins on the other hand absorb best at 280 nm and organic...you can compare the fluorescence of your sample against this curve to quantify your DNA prep. Though this...this method doesn’t tell you about contaminating proteins or chaotropic salts. Capillary electrophoresis...read with a standard ELISA reader if no other instruments are available. Some last reminders about quantifying... -
Using CRISPR/Cas9 to Edit Disease Out of the Genome
TypeBlog PostPublishedJune 23, 2022, 1:15 p.m....in monoallelic diseases In 2013, a team led by Jinsong Li from the Chinese Academy of Sciences found that...independent team led by Hans Clevers at Hubrecht Institute in The Netherlands used the CRISPR/Cas9 genome...handle on how cleavage sites are repaired.” Those insights will be critical for scientists’ ability to influence...addition, the current work in developing anti-CRISPR proteins aid the limitation of these mutations, to control...s Disease.” Journal of Alzheimer's disease & Parkinsonism. https://doi.org/10.4172/2161-0460.1000439 6... -
Photosensitizer Induced Cell Ablation with FAP-TAP MG-2I-dL5**
TypeBlog PostPublishedSept. 19, 2017, 1:20 p.m....Wang, Y., Missinato, M. A., Onuoha, E., Perkins, L. A., Watkins, S. C., . . . Bruchez, M. P. (2016). A ...you can do just that. Photosensitizing dyes and proteins have been around for awhile (check out this review...produced by photosensitizers inactivate target proteins or cells while having little effect on neighboring...suggests that it could be used with these fluorescent proteins. He et al also compared MG-2I-dL5** head-to-head... -
Pushing the Limits of DNA Assembly
TypeBlog PostPublishedSept. 6, 2017, 1:01 p.m....sections, while still creating scarless plasmid inserts. Want to Learn More about DNA Assembly? Check...polymerase used in the amplification. This tool allows instant updates as fragments are added, deleted, or reordered...(sgRNA). Using only one single-stranded oligo, instead of the more typical two oligos, is more cost effective...linearized plasmid with a synthetic oligo that contains a region of homology and any desired edits to ...amplified with overlapping regions of homology and inserted into a linearized cassette, ready for transformation... -
Hot Plasmids - January 2023
TypeBlog PostPublishedJan. 31, 2023, 2:15 p.m....second GFP antibody? Similarities: Generated against full length GFP and converted to recombinant form...Isotype IgG2a Differences: Distinct variable domains that recognize different epitopes on GFP Monoclonal...light over the last decade, but the fluorescent proteins (FPs) necessary for many of these applications...consider hfYFP and LSSmGFP. Fig. 4: hfYFP retains folding and fluorescence in high molarity GdnHCl...Campbell, B.C., et al. Chemically stable fluorescent proteins for advanced microscopy. Nat Methods 19, 1612–... -
Four Factors that Differentiate the Stem Cell Field
TypeBlog PostPublishedSept. 28, 2016, 2:35 p.m....differentiated cells simply by expressing four proteins Oct4, Sox2, Klf4, and cMyc (the so called OSKM...Yamanaka's work has been profound, and this is a rare instance in which "profound" seems like an understatement...degeneration; Laurenz Studer discussed his work with Parkinson's Disease; Hideyuki Okano presented ground-breaking...clinical trials for ALS. I have an uncle with Parkinson's, a half-brother who is a quadriplegic due to ... Eric J. Perkins... -
5 Great Apps for Lab Life
TypeBlog PostPublishedSept. 22, 2016, 2:30 p.m....searching the interwebs for lab resources, and instead conveniently provide them on your smart phone. ...holds the Gibco Cell culture manual and basic instructional videos. The search function in the manual just...that require restriction enzymes or PCR. The app contains a double digest tool that is only helpful for ...for NEB restriction enzymes since the instructions use NEB buffers. Nonetheless, if you are setting up a...draw an amino acid or peptide for some reason. It contains all twenty amino acids plus selenocysteine and... -
OCaMP: A New Calcium Indicator for Neural Imaging
TypeBlog PostPublishedAug. 19, 2025, 1:15 p.m.... at the Hotchkiss Brain Institute. He previously worked at The Allen Institute for Neural Dynamics and...tool in neuroscience research. These fluorescent proteins light up in response to calcium, which is more...spectrum (Figure 1). The GCaMP series (e.g., jGCaMP8) remains the gold standard for green fluorescence, while...Additional Resources on addgene.org Fluorescent Proteins: Biosensor Plasmids In-Stock AAV Vectors: Biosensors... -
Q&A with Karen Katz and Carl Paratore, the Newest Members of the Addgene Board of Directors
TypeBlog PostPublishedFeb. 19, 2019, 1:36 p.m....certified health insurance executive (CHIE). He is a member of both the Internal Institute of Auditors and... Directors brings complementary experience and insight to the now 10 member board at Addgene. Let’s meet...at both MetLife & Ernst & Young, LLP, auditing insurance, finance and high-tech industries. He has earned...Addgene blog Learn more about about Addgene in our "Inside Addgene" blog series Subscribe to the Addgene blog... -
A Quick Guide to a Career in Software Product Management
TypeBlog PostPublishedNov. 24, 2020, 2:15 p.m.... thanks to our guest blogger Rachel Rubinstein! Rachel Rubinstein is currently a Field-based Product Manager...post was contributed by guest blogger, Rachel Rubinstein, a field-based software product manager at Oxford...your network to your advantage As the profession gains more traction in the sciences, I believe that more...customer in mind. You can follow her on twitter @rgrubinstein. Additional resources on the Addgene blog Discover... -
Clear and Accessible Writing
TypeBlog PostPublishedFeb. 15, 2024, 2:15 p.m.... may feel it’s overwhelming or challenging. So instead, let’s focus on practicing with a short and accessible...information? This can be physical or digital. For instance, you would present information differently in ...image on a protocol page on our website, as an instructive step-by-step guide. Finally, when writing your...after centrifugation and the syringe needle is inserted bevel-up at the interface between the 40% and ...captured at the 40% layer. To collect purified AAV, insert a syringe at the interface of the 40% layer containing... -
Viral Vectors 101: Biosensors
TypeBlog PostPublishedJuly 6, 2023, 1 p.m....readouts for levels of different chemicals, hormones, proteins, etc., that you are interested in. The most obvious...could be a microscope, plate reader, or any other instrument which measured GFP levels and converted that ...includes tension monitors associated with specific proteins of interest such as actinin. When tension is high....2019.00474 Resources on Addgene.org Fluorescent Proteins: Biosensors Viral Service: Biosensor AAV Viral...Epigenetics with a FRET Based Biosensor Fluorescent Proteins 101: Fluorescent Biosensors ... -
AAV Q&A with Tim Miles
TypeBlog PostPublishedMay 17, 2022, 1:15 p.m....AAV? A: One potential strategy is to use split-inteins to mediate trans-protein splicing. Note, however...possible to make an efficient AAV prep for a 4.2 kB insert with the CMV promoter using the AAV-BR1 capsid?...bringing total genome length to ~4.7kB. If your insert alone is 4.2 kB and CMV promoter+enhancer is 0.5...experiments using PHP.eB in the brain give some insight into codelivery efficiency at different doses. ...tropism to other serotypes, but reproducible success remains elusive. There has also been a lot of effort in... -
Seven Tips for Using LinkedIn as a Scientist
TypeBlog PostPublishedOct. 1, 2019, 1:21 p.m....discussions. Pro tip: Instead of connecting with someone you don’t really know, follow instead of connecting....information available. When I am speaking at a new institution, for example, I will often see a whole bunch ... “a postdoc from this Uni viewed your profile” instead of “Joanne Kamens, PhD viewed your profile.” The...option for sending LinkedIn messages to Click Send instead of Press Enter to Send. That way you can type a...