...conformational changes lead to an increase in fluorescence intensity from the inserted GFP. After characterizing iGluSnFR...by glutamate, and tweaking it to improve its responsiveness, they showed that iGluSnFR could detect neuronal...and produces stronger fluorescent signals upon sensing glutamate in mouse and ferret models. Furthermore...recently made iGluSnFR3 variants that are more sensitive and better localized for synaptic glutamate response... and produce stronger fluorescent signals upon sensing glutamate using both 1-photon and 2-photon imaging...

...can be engineered to be cell type specific. Once inside the host cell, the virus unleashes its cargo – ...signals which can traffic their viral particles inside the nucleus, without the need for envelope disassembly...viruses do. They are generally used to achieve transient, high expression of the relevant cargo without...Capacity (>8kb) High Transduction Efficiency Transient Gene Expression High Immunogenecity AAV ...Immunogenicity Infects Non-Dividing Cells Transient Gene Expression Low Tropism for Mouse Cells and...

...personalized steps like “check drawers for light-sensitive reagents” or “re-sterilize now.” Have a physical...Soon you’ll figure out what prompts you need to consistently and accurately run a protocol.   Figure...experiment than it is to rerun an experiment, so consider your prep an investment in accuracy.  Bench setup...hasn't. Developing simple habits like these can consistently reduce errors as you work, and the possibilities... to a protocol if you’re in a hurry, but let’s consider it this way: When you’re stressed or rushing, ...

...DNA packed into our chromosomes is very much 3-dimensional, as you can see by this post’s chromatin illustrations...PIs, articles, genes, and blog posts are not comprehensive, so just because you do not see a name or gene...the gene you’re looking for on this list, please consider searching directly on our website. In-exhaustive...Reporter-nanobody fusions (RANbodies) as versatile, small, sensitive immunohistochemical reagents. Yamagata M, Sanes...chromatin and the epigenome. If you have been considering some new techniques, the posts below may steer...

.... The next insight was to assemble a synthetic transcription activation complex consisting of multiple...transcriptional activation of endogenous genes and it consists of three components: 1. A nucleolytically inactive...-mediated screens exhibited a high degree of consistency and validation with >80% effectively enriched...optimize humans for CRISPRi and CRISPRa.” Before considering the use of pooled libraries in your experiments...

...They all found the support of the group to be intrinsic to their success. Their stories made it clear ...programs. The groups in these programs typically consist of 1-2 mentors and 3-5 mentees who meet together... of people who are all interested in making a transition. Schedule your first meeting and get organized...in a rotation. The leader for that meeting is responsible for choosing a topic and leading the discussion... your successes and support each other as you transition to the next stage in your careers! Stay in touch...

...secondary considerations to validation, application specificity, and experimental considerations - no other...has been validated in your specific application. Consider also that there may be variations within an application... your needs.    Multiplex experiments Finally, consider whether you will be using other antibodies in ...from a vendor that you use frequently, with a responsive and knowledgeable technical support team, is ...

...channelrhodopsin used as a sensitive fluorescent voltage indicator with near-complete desensitization in continuous... the imbalance of electrical charge between the inside and the outside of the cell. This imbalance is ...Q239 - A GEVI based on the fusion of the voltage sensing domain of a Ciona intestinalis voltage sensor and...anion channel. It is triggered by a short low-intensity blue light pulse and stays open for a longer duration...

...proteins: a relatively pH-insensitive RFP variant, DsRed.T3 and a pH-sensitive GFP variant, super ecliptic...Rosella is a pH-sensitive fluorescent biosensor that was recently deposited with Addgene by Dr. Mark ... and recycling. The key to Rosella’s autophagy-sensing abilities is that its fluorescence emission spectra...regardless of its localization in the cell. SEP’s pH sensitivity means Rosella will fluoresce green unless it’...

...spectral properties of the fluorophores you are considering to make sure that they’ll be compatible with ...antibody conjugates (especially if you start to consider clinical applications). But there are a few other... at least mention. Oligonucleotides allow for sensitive detection of low abundance targets with significant...mechanisms. Most take advantage of chemical groups intrinsically present on antibodies, such as the primary amine...can conjugate an antibody yourself allows you to consider a broader selection of antibodies for your experiments...

... learned over the years with the hope that our insights can help steer you in the right direction when... of this is gospel, but we think it's worth consideration. Everything will go wrong Whenever an order ...easily as needed. In this case, the physical considerations of the laboratory make object composition a...technical details of the problem you're solving, but consistency is key. Some namespace collisions you may encounter...treating these pieces of data in ways that are inconsistent with how they are being treated in the lab, ...

...given problem, Empowers the writer, And offers insight into a career as a Professor. Step 1: Your ...sections in a NIH RFA, for example, and tips to consider while writing each section. Research challenge...time to review the literature and offer educated insight into what the desirable features of future technologies...deem your proposal successful. Typical proposals consist of 3 to 5 aims, with secondary milestones to meet... the application. Make sure to invest time and consider the reviewer's comments. Whether the proposal ...

...virion is a non-enveloped particle of ~100 nm, consisting of a capsid surrounding an inner core which contains...adenovirus: the virion is ~ 100nm in size, and consists of an icosahedral capsid made of 3 types of proteins... for creating adenoviral vector constructs. It consists of 2 plasmids: a shuttle vector (in which the ...purification of rAdV uses cesium chloride (CsCl) density gradients combined with ultracentrifugation to ... rAdV genomes/mL) can be measured by: Optical density: particle concentration is measured by looking ...

...Interactions Difficult to detect weak and/or transient PPIs. Many PPI identification techniques rely ...often prevents the identification of weak and/or transient interacting proteins. No spatiotemporal control...mass spec analysis. Biotinylated proteins are considered possible interaction partners if they are enriched...BioID, is also useful for identifying weak and/or transient protein-protein interactions since the biotin ...cell lysis and lysate fractionation. Important considerations for Split-BioID It’s important to remember ...

...some reliable controls. 2. Viral stocks may be sensitive to freeze thaw cycles. Depending on the researcher...ultracentrifugation of the viral stock, followed by resuspension of the collected particles in a smaller volume...Davis et al., 2002). However, polybrene is highly sensitive to freeze-thaw cycles and should be stored in ...enhance transduction efficiency in cells that are sensitive to the toxicity of polybrene (e.g., hematopoietic...PMID: 12050007. 7. Stockschläder, M., et al. "Expansion and fibronectin-enhanced retroviral transduction...

...III), each responsible for transcribing different types of RNA: Polymerase I is responsible for ribosomal... Rho-independent termination is also known as intrinsic termination, and it relies on the formation of...ribosomal RNA, Polymerase II is responsible for mRNA and miRNAs, and Polymerase III transcribes tRNA and other...polymerase-associated proteins, CPSF and CstF, which are responsible for recruiting the cleavage and polyadenylation...

...injections are most often used when injecting a cell suspension to create xenograft models. Restraint: back scruffing...sounds very disruptive, but surprisingly, it is considered minimally invasive. Restraint: anesthesia Location...common. Due to the intubation, this procedure is considered quite invasive. Restraint: anesthesia Location...introduction to common injection routes in mice! When considering what method you’d like to use, remember that ...animals: routes of administration and factors to consider. PubMed, 50(5), 600–613. https://pubmed.ncbi.nlm.nih.gov...

...to their high density, which can be collected afterwards. You can then remove the density gradient material...certain in vitro and in vivo applications, you can consider using unpurified AAV from the supernatant (Goodwin...cells and supernatant then need to be purified by density gradient centrifugation (e.g., iodixanol) using...assay for adeno-associated virus vectors with sensitivity sufficient to detect single infectious events...

...this temperature and can bind to the primers.  Extension- The temperature is then raised slightly to Taq...  This process of denaturation, annealing and extension is repeated 25-35 times to exponentially replicate...droplet PCR or ddPCR is a method that provides ultrasensitive and absolute nucleotide concentration, unlike...more complicated to develop and is often less sensitive than PCRs that use a single pair of primers, like...Diagnostics NGS library prep QC ddPCR Determine ultrasensitive and absolute nucleotide concentration of starting...

...because they are inside the virus itself. While these impurities are generally considered innocuous for ...helper plasmids or the cell genome can be packaged inside the vectors during production. These impurities...NGS at Addgene to sequence all the DNA packaged inside the viral particles (Figure 1). Briefly, packaged...contamination is a good start, but to determine the sensitivity of our method we wanted to see how low we could...