...luminescent opsins: fusion proteins of a light-emitting luciferase and a light-sensing optogenetic element, making...oxidizes its substrate (luciferin), it generates light that activates the opsin to either excite or inhibit...

... for each specific antibody in order to generate high quality preparations. Following production, all ...transfected with the plasmid(s) encoding the heavy and light chains. Antibodies are expressed for 7 days and ...amplified with primers targeting the heavy chain and/or light chain variable regions. The PCR product is purified...

...joining fragment comprising a κ light chain constant domain (C L ) and the κ light chain polyA tail sequences... for light chain expression, and an ER signal/leader sequence (L) for translocation of the light chain...in a new window) Durocher Y, et al. 2002. High-level and high-throughput recombinant protein production... R-mAbs are secreted into the culture media. For high level expression we recommend using the HEK-2936E...Link opens in a new window) . The schematic to the right shows the separate elements of the R-mAb expression...expression plasmid involved in coexpression of light (green) and heavy (blue) chains as driven by two CMV promoters...

...Vector Core) and Addgene have partnered to provide high-quality AAV vectors to the academic research community...Deisseroth AV-1-36857M 100038-AAV1 pENN.AAV.hSynap.Flex.ArcLightDco.WPRE.SV40 Vincent Pieribone AV-1-36857P...36857P 100037-AAV1 pAAV.hSynap.ArcLightD.WPRE.SV40 Vincent Pieribone AV-1-51473 51473-AAV1 pAAV-hSyn1-mRuby2...James M. Wilson AV-1-PV2699 100030-AAV1 pAAV.CBA.loxP.ArcLightD.2A.nlsmCherry.loxP.WPRE.SV40 Lawrence Cohen...

... are able to recognize their targets with fairly high specificity. Fifteen years of research has shown...TALe research has been swift. The incentives are high. Since ZF targets are confined to sequences composed...on specificity is on par with ZF arrays, if not slightly lower. Figure 2: Simplified representation of ...

...and scFvs for neuroscience research created with high-volume hybridoma sequencing on the NeuroMabSeq platform...dissemination to the research community. This collection highlights plasmids created with support from the BRAIN ...

...based on using a thin plane (sheet) of light generated by sunlight to observe single gold particles with...V= ventral Developments in light sheet fluorescence microscopy Modern light sheet fluorescence microscopy...out-of-focus reflected light by using a pinhole.  However, the excitation light excites all of the fluorophores..., there is a drop in image brightness with increasing depth caused by light scattering and absorption....of this century has seen some major advances in light microscopy, particularly related to the neurosciences....  One of these advances in microscopy has been light sheet fluorescence microscopy (LSFM). The underlying...a fluorescent sample and captured the reflected light using a different objective lens oriented perpendicular...

...pAAV-EF1a-iCreV EF1a light-inducible recombinase iCreV Zeng 140136 pAAV-EF1a-iDreV EF1a light-inducible recombinase...recombinase iDreV Zeng 140137 pAAV-EF1a-iFlpV EF1a light-inducible recombinase iFlpV Zeng 164450 CN1851-rAAV-hI56i-minBglobin-iCre...

...hundreds of in-stock preps that are ready to ship right away, as well as our Packaged on Request service...expressing Rep2 and B1 capsid (backbone may yield higher titers than pAR-B1) Sena-Esteves 78503 pAR-B1(1986.1...systemic delivery Maguire 198014 pCMV-AAV2cap AAV2 For high-level expression of the AAV2 cap gene, suitable ...

... clones for loss of function. This is due to the high frequency of indels formed on non-deleted alleles...cells in order to enrich for cells that received high levels of the CRISPR/Cas9 constructs. Plate sorted...the CRISPR-Cas9 system. Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F. Nature Protocols...

...data on the assay methods. You might find that while the website highlights a variation of your application...protein or protein region highly conserved across species? If the proteins are highly homologous between your...

...experiments require higher gene expression and therefore higher titer than others. High titers are often ...vector to such a high titer when looking for effective gene expression. How do you achieve high titers in the...good ones into a cell to produce expression. The higher the titer, the better. AAV transport and transgene...transgene expression It’s clear that we need high titers, as contact between the cell and any individual AAV...the cell may not always be enough and therefore higher titer will be required. For example, retrograde...over an even larger area, thus necessitating even higher retrograde AAV titers for successful transduction...That means that once you drop below a certain (very high) threshold of viral particles per cell, you may ...

...the pressure to publish in high-impact journals, and the requirement for a high grant success rate. At this... this point, candidates are highly experienced but often have to start pursuing other options. While some...technical support specialist for 4 years. Read on for insights from my time in technical support including tips..., and resolve complaints about products. As you might expect, different companies produce different products... dialogue. This will give you a feeling of what might be involved in such a role. In addition, it can ...individual stories and get (sometimes refreshing) insights into how other scientists think and work. Most...to enhance the OD signals and then complain about high background. In many instances, variations of the...

...for many years (at a very low salary I might add). You have a right and responsibility to choose a lab where...Why is choosing the right lab such a big deal? It’s actually something you CAN choose and it will make...difference for your future career and life. You might see a lab head as choosing you, but in reality, ...communication before making this important decision. I’ll highlight some of these questions in the sections below....you are good at asking for and getting help, size might not be a big factor for you. Know who's boss You...considering a career at a small, teaching college you might want to consider less expensive, portable areas ...ownership" and author order PI lets lab members fight it out PI intervenes if there are interpersonal...

...antibodies have higher affinity and specificity than mouse monoclonal antibodies. Antibodies with higher affinities...monoclonal antibodies contain a single clone they have higher lot to lot specificity and tend towards more reproducible...When using a monoclonal antibody be sure to use a highly specific secondary antibody, rather than a broadly...at very low levels or your application requires a high degree of affinity, then species choice could very...may be the better choice for assays that require high affinity binding, such as immunoprecipitations. ...stain a target protein in mouse tissues, causes a high degree of background staining. This occurs because...antibody isotypes matter and helps you choose the right antibody for your application!  References and ...

...fluorescence when exposed to 500 nm light.  Under 400 nm cyan light, Dronpa dimers dissociate, Dronpa goes...minute pulse of 500 nm light (turn on psRaf1) followed by 3 seconds of 400 nm light (turn off psRaf1). After...and use MEK1 as a model kinase. In this system, light is used to turn “on” and “off” MEK1 activity by ...is called psMEK1. When exposed to 500 nm violet light, the Dronpa dimers interact and lock MEK1 in an ... an inhibitor of choice, exposed to 500 nm cyan light to activate psMEK1, and the localization of ERK ...these same cells by 24-48 hours exposure to blue light also induced tail swelling in ~75% of worms (see...expressed in a tissue or organism that blue and violet light can penetrate. How can I try using photoswitchable...

... short generation times and high fecundity, make zebrafish ideal for high throughput studies of optogenetics...Optogenetics, the use of light sensitive proteins (opsins) to manipulate cell activity, enables researchers... studies because it is non-invasive and offers a high degree of control over time and space. Zebrafish...imaging with fluorescent tags and activation of light sensing proteins. Furthermore, during the first ...Optogenetics functions through opsins which are light sensitive proteins. Opsins used in optogenetic studies...particles to cross the cell membrane when exposed to light. Neuronal firing depends on these charged particles...of the differing peak absorption wavelengths and light intensity requirements of different opsins, studies...

...solution for discriminating between neurons for light microscopy-based neuronal reconstruction. To achieve...et al., 2007; Cai et al., 2013). However, the brightness of Brainbow was inherently low. This is because...at ~2 copies per cell per color to achieve the highest color variations. Adjust the plasmid concentrations... to express a minimal amount of tTA to achieve highest expression levels. Clearing large tissues - Fluorescent...anatomical aspects of sensory circuit development using light microscopy. Lab website: https://dn-en.med.kyushu-u.ac.jp...clearing and index matching of tissue samples for High-resolution fluorescence imaging using SeeDB2." Bio-Protocol...Sakaguchi, Richi, Marcus N. Leiwe, and Takeshi Imai. "Bright multicolor labeling of neuronal circuits with fluorescent...

...excitation light less efficiently. Similarly, decreasing the power of the excitation light reduces the...series described a practical approach to selecting a bright fluorescent protein. In the second post of this...destruction of a fluorophore under the influence of light. Any fluorescent molecule will photobleach at some...fluorescent proteins are excited with very intense light (for a brief time), whereas in widefield imaging...imaging fluorophores are excited with relatively low light levels (for a prolonged time). Other illumination...that, in experiments that measure photostability, high excitation powers are often used to reduce the length...linearly related to power, the conclusions reached at high power may not translate to real applications in ...

...dark state and highest fold induction at 31X. By using a slit pattern during blue light exposure (470nm...proteins N713-pMag and nMagHigh1-C714, this new system had both low background and high fold-induction of ...reversibly activated by turning blue light on and off. As one might expect from the modular nature of Cas9...fused with high-affinity interacting proteins Coh2 and DocS, respectively. The use of far-red light is of ...particular interest since blue light has limited applications in vivo. UV and blue light is readily absorbed and...tissues. Additionally, high intensity exposure can cause DNA damage. Far-red light, however, can penetrate...protein. Stop light! Photoactive anti-CRISPRs halt Cas9 activity Anti-CRISPR proteins are a highly diverse ...