...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 ...

...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...

... 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...

... 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...

...resources. There are three types of cognitive load: intrinsic cognitive load is the effort associated with a...show frustration if they're incorrect; take it instead as valuable feedback.  Listen. Think about the ...strategy. Don’t assume it’s the audience’s fault; instead look for ways to improve how you're communicating...If something I was communicating didn’t click, instead of blaming the audience, I would think about how... wasn't. Maybe I needed to employ a visual aid instead of describing something verbally. Sometimes a group...

...for every assay. Antibodies that are generated against a linear antigen tend to work very well for applications...application. Take immunohistochemistry (IHC), for instance. While an antibody may be validated for IHC, not...region highly conserved across species? If the proteins are highly homologous between your target species... each isotype or species to easily discern the proteins in your assay.  Narrowing the field After working...compare the staining patterns of multiple antibodies against the same target; tagged protein strategies that...

...awards to early career scientists using fluorescent proteins in their research. These awards help early career...biosensor fuses MPST to roGFP2. Because roGFP2 contains two cysteines near the chromophore that are susceptible... to better understand the processes that happen inside the cell,” says Pedre Pérez. Figure 2: ...changes following opioid withdrawal, providing insight into new methods for treating patients in recovery...indicator which had improved performance in live mouse brains. Currently, she is using the improved voltage indicator...

...postdoctoral researcher at the Noble Research Institute, co-instructed the ASPB workshop and authored a three-part...well received, and then when the team started brainstorming ideas for the next workshop, they realized that...in the group was able to make it physically to instruct the workshop at the conference, but because each...of the scientific community. Lessons learned Instead of continuously talking about a problem and potential...

...comprised of two tandem fluorescent proteins: a relatively pH-insensitive RFP variant, DsRed.T3 and a pH-sensitive...the degradation rate of radio-labeled long-lived proteins as a proxy for autophagy. Cells are cultured with...washout period removes radio-labeled short-lived proteins primarily via proteasome degradation. Autophagy... to giving more fine-grained detail about what proteins or organelles are being degraded) and it is slow...

...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...

...destination vector + insert may spontaneously religate, this transient construct retains functional Type IIS...in the final construct. The destination vector contains sites with complementary overhangs that direct...complete with helpful tips and tricks, see The Sainsbury Lab website or Engler & Marillonet. Golden Gate...technology has adapted Golden Gate cloning for inserting the appropriate oligonucleotides specifying a ...

...mutation in the evolved TadA domain. Because ABE7.10 contains a heterodimeric fusion of wild-type TadA and TadA7.10... the evolved TadA region. The ABE8.x-m version contains only the evolved TadA and is ~500 base-pair smaller...evaluate further.  The creation of accidental insertions and deletions (indel) by the catalytically impaired...more than 90% compared to ABE7.10. Choosing Cas9 proteins to increase the targeting scope of adenine base... Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes NG-Cas9 (PAM: NG) to create NG-ABE8...

...challenge is finding relevant papers. The latest installment of the Early Career Researcher Toolbox will highlight...search that’s constantly updated. This Twitter list contains >300 Twitter literature bots, including a bot ...AI-backed literature search developed by the Allen Institute for Artificial Intelligence. Semantic Scholar ...Initiative. Meta is not a traditional search engine. Instead, you create your own research feed by entering ... papers are published, similar to Facebook or Instagram. Key features Setting up a personalized feed is...

...distinct proteins and result in unclear transcriptional consequences (Sprujit 2013). Enzymatic domains tethered...are a number of drawbacks to this approach: TET proteins are not demethylases; they are dioxygenases that..., Charlton et al. 2020). Most importantly, TET proteins affect transcription independent of their enzymatic...overlap the CpG of interest. Having the CpG directly inside the gRNA sequence is a great guarantee that DNMT1...

...secondary antibodies instead (B).   Since a polyclonal primary antibody contains a mix of different...antibodies used for detection bind to non-target immunoglobulins naturally present in the tissue sample. There...antibody fragments to block the endogenous immunoglobulins and reduce the background staining but they...Biological systems tend to be complex with a variety of proteins interacting. Experiments tend to be equally complex...

... they bind to a class of Argonaute proteins. The Argonaute proteins remove one strand of the siRNA and...cDNA), and then use PCR and molecular cloning to insert the cDNA into the desired plasmid backbone. The...the process is a little more complicated. These strains are more picky eaters and refuse to eat liver paste...Sánchez lab protocol also provides step-by-step instructions for the “the bravest and more curious” students...

...first make your poster. While a poster generally contains the same sections as a primary research article...in your poster. This is a section where you can insert your references and contact information, and it...conclusions, and acknowledgements sections are inserted. The advantage of this style is that it will display... a poster vertically rather than horizontally. Instead of displaying your information as columns, you ... poster, written and graphical information are inserted into larger circles or ovals, that are arranged...

...use a standard strain, like DH5α, grown at 30 °C instead of 37 °C. If you’re unsure about the integrity ...prefer to store freshly harvested virus at 4 °C instead of storing it briefly at -80 °C. If troubleshooting...option may be to avoid freezing altogether, and instead to use the virus immediately upon harvesting. Interestingly...appear as a white pellet. The pellet can then be rinsed and resuspended in cold sterile PBS.  Alternatively...of ensuring viral production is to perform PCR against components of the viral vector using the harvested...

...Director of the National Human Genome Research Institute (NHGRI) of the NIH, started off with the first...will lead to advances in human health. It was inspiring to hear how the genome project has already had...enzymatic machinery (i.e. RNA replication with no proteins). The chemistry is complex and this work requires...desribed how P. furiosus has systems to defend against both DNA and RNA viruses - 2 cas systems regulate...system. His lab is also exploring the use of cas9 proteins from different species for genome engineering....

...—an optical insulator called a ferrule (below). The fiber optic cable/ferrule is inserted into the target...stimulation guarantees that the majority of the opsins will receive sufficient quantities of photons to...although you may be able to purchase one pre-made instead). Laser stimulation         In the video above...: closed/light receptive, open, closed/light insensitive. If you deliver a continuous stream of photons...