...not recommended that lentiviral supernatants be subjected to multiple freeze-thaw cycles. This protocol ...

...blog post shows. To avoid these issues, Subach et al. and Tsuboi et al. developed monomeric FP timers with...monomeric FP timers (mK-GO from Tsuboi et al. and the FT series from Subach et al.) were derived from previously...membrane protein, and following its subcellular localization overtime (Subach et al.) Monitor protein expression...tetramer with a propensity to aggregate in cells (Tsuboi et al.), limiting its use. Nonetheless, its creators...fluorescent proteins mKO and mCherry, respectively. While Tsuboi et al developed mK-GO somewhat serendipitously ... while attempting to enhance mKO in other ways, Subach et al made a concerted effort to develop their ...see Table 1 for emission and absorption spectra). Subach et al’s directed mutagenesis also produced the ...

... out Cas3. Type I is further subdivided into subtypes A–G. These subtypes have similar compositions and...immunity. Within type III, there are six subtypes, A–F. All subtypes use Cas10 as the primary effector complex...into a fairly simple hierarchy: class to type to subtype to variant. Despite the seemingly complicated steps...types are based on main function and composition; subtypes and variants are typically based on species of...There are currently two classes, six types, 33 subtypes, and 17 variants. Figure 1: Overview of...hierarchy. The hierarchy flows from Class to type to subtype to variant (shown in italics). Created with BioRender.com...Figure 2: Schematic of a Cascade complex (blue subunits) with a guide RNA (green and orange) bound to ...

...different subclasses, but there are also secondary antibodies that are specific to each subclass. Approximately...mouse mAbs are IgG1 subclass (Manning et al. 2012), which limits the use of subclass specific secondary...backbone to generate R-mAbs of different subclasses, including subclasses that were not produced by the original...original hybridoma. By using subclass specific secondary antibodies and subclassed switched R-mAbs, multiple... by some of the hybridomas and, as a result, subsequently cloned into the expression constructs. The Trimmer...engineered. Like humans, mice express multiple subclasses of IgG antibody. Broad range secondary antibodies...  Fig. 2. Multiplex immunolabeling with subclass-switched recombinant antibodies in adult rat brain...

...unique antibodies that bind discrete subunits or particular subunit pairs. Some of the antibodies in the... one of 18 alpha and one of eight beta subunits. Both subunits have short intracellular ends and large...region displayed by a subset of the RGD-binding integrins containing the αV subunit. Using these affinity...discovered by yeast display reveal αV subunit pairing preferences with β subunits (p. 2024.01.26.577394). bioRxiv...well-known motif serves as a recognition site for one subfamily of integrins, the RGD-binding integrins. Six of...remarkable specificity for their targeted integrin subtype, even when in competition with other similar integrins...antibodies to show the binding preferences of the αV subunit and its β partners. Of interest, we demonstrated...

...not recommended that lentiviral supernatants be subjected to multiple freeze-thaw cycles. Procedure Before...

...three subunits: alpha (Gα), beta (Gβ), and gamma (Gγ). G proteins are divided into four subtypes, based...associate with certain subtypes, although most are able to signal through more than one subtype. How GPCR signaling...disassociation of the Gα subunit from the Gβγ dimer. The Gα and Gβγ subunits then induce or inhibit intracellular...based on their Gα subunit: Gαs, Gαi/o, Gαq/11, and Gα12/13. Gβ and Gγ are closely associated and function...change in the GPCR causing the activation of the Gα subunit. The activated Gα exchanges bound GDP with GTP,...inactive heterotrimeric G protein complex and subsequently another round of signaling.    Addgene Tools...plasmids encoding 14 alpha, 2 beta, and 4 gamma subunits of the heterotrimeric G protein complex that can...

...influence your choice of journals to submit your work to. Before submitting your paper to your preferred  ...approve your paper. Rush your submission or revision. A sloppy submission, missing files or a manuscript...scientific community, or are you writing for a specific subgroup of researchers in your area of expertise? Sometimes...who wishes to read it, without them needing a subscription to the journal. An increasing number of funding...a reliable source of information, it is best to submit it to a journal that uses a peer review system....if you don’t. Add a clear cover letter to your submission, briefly summarizing the conclusions of your ... the Author Guidelines of the journal you are submitting to and make sure your manuscript fits the house...

...community submissions for Addgene and CLOVER AAV experiments - and we've updated our submission process ...say, should!) submit your data to the Data Hub! The process is easy with our new submission form - and no...step-by-step submission guide or email [email protected] to learn more. A great submission includes: ... cited by others?)  Ready to submit?  Fantastic! The next 25 submissions will receive a 20% off coupon...recording to see two reports presented by the submitting scientists, learn more about the AAV Data Hub...given by Connie Cepko and Ian Wickersham.     Submitting to the Data Hub If you've ever validated an Addgene...

...0.02–0.05 ml Skill required: beginner Subcutaneous Subcutaneous (SQ or SC) injections are given between...method, subcutaneous injections take some practice to get the coordination down. Subcutaneous injections...the mice, and restraint method. Many different substances can be injected, including cells, drugs, DNA,.... IV injections are used for a broad range of substances, as it circulate what is injected throughout ...technique. Intrathecal injections are used when the substance being administered must be delivered to the central...Intratracheal instillation is used to deliver substances directly to the lungs. It is an advanced technique...invasive, and only requires a pipette. Since the substances are administered through the nose, they also ...

...the standard curve is y = 14725x + 3748.4, then substitute 18179.442 for y and solve for x. 18179.442 = ...

...centrifuge with the mark facing outward in all subsequent centrifugation step Pro-Tip Resin will appear...

... deliver the substrate; however, secreted luciferase or the use of alternative substrate reagents can ...reaction requiring luciferin, ATP and oxygen as substrates. Production of photons by this bioluminescent...of Photon Generation Slower Faster Cofactors/Substrates Required Not Required Signal Strength Lower.../Phototoxicity Not susceptible Susceptible Subcellular Imaging Improving Well-established High-throughput...identified in various species with differing kinetics, substrate requirements and photon emission wavelengths. ... upon catalyzing a chemical reaction with the substrate coelenterazine and is likely the second most commonly...crustaceans. Many of these luciferases have been subjected to further engineering to improve upon the properties...

...neuronal subtypes was not available until now. In order to target specific neuronal subtypes such as inhibitory...recently deposited 2 Bacillus subtilis libraries with Addgene – the B. subtilis Single Gene Deletion Library—Kanamycin...Every quarter we highlight a subset of the new plasmids in the repository through our hot plasmids articles...hot plasmids segment on the Addgene Podcast. Subscribe to the Addgene blog to get all the latest hot ...containing high fidelity Cas9 variant HF-Cas9. They subsequently tested the original BE3 and HF-BE3 using a gRNA...the podcast Segement on the pPSU Plasmids B. subtilis knockout and knockdown libraries The Gross Lab... Essential gene Knockdown library. Although B. subtilis has a fully sequenced genome, many applications...

...are not subject to photobleaching/phototoxicity, they successfully imaged multiple subcellular compartments...luminescent substrate is required for imaging, and it is important to verify that this substrate does not...fused to an FP; when supplied with a luciferase substrate, the luciferase transfers energy to the FP, resulting...excitation light source. Instead, a luciferase substrate like coelenterazine is supplied in the culture... which is limited by depletion of luminescent substrate, they synthesized diacetyl coelenterazine-h. Unlike...coelenterazine-h, producing a constant supply of substrate. Using this compound to excite the Nano-lanterns...brightest characterized luciferase, NanoLuc, and its substrate furimazine. In addition to the previously described...

...ligate many undesirable structures, including substrates containing gaps or DNA base pair mismatches (...such as Taq DNA Ligase, which only reacts with substrates containing no gaps, and will not join any fragments... sequence is present, the probes will ligate. Subsequent cycling through rounds of melting and annealing...generally prefer fully Watson-Crick base-paired dsDNA substrates to those containing one or more mismatches. However...pyrimidine mismatches are typically worse ligation substrates than pyrimidine:purine mismatches. Helix stability...crystal structures of several DNA ligases bound to substrate (23, 24). Thermostable DNA ligases, including ...naturally able to discriminate against ligating substrates containing base pair mismatches (i.e., are "higher...

... have subscriptions to the specialized journals in your field? Post a preprint before you submit to a ...article, presenting a conference poster or talk, or submitting patent or grant applications; to informal settings...especially the increasingly growing time from submission to publication. It takes many months, often more...the possibly repeated rounds of rejection and resubmission to other journals. Preprints allow authors to...admittedly, somewhat meaningless) practice of listing “Submitted” articles in their CV or grant applications. A...preprint at the same time as, or slightly before, submitting the manuscript to a journal for formal peer review...opportunity to strengthen the manuscript prior to submission. Preprints at Addgene Preprints are all about...

...cytosine-to-thymine substitutions in target sequences (Banno et al., 2018). These substitutions were limited ...enable C-to-T and A-to-G base substitutions respectively. Substitutions seem most efficient when using...time needed for a single round of editing, and a subsequent curing protocol that requires only 2-3 hours ...days, and the pTargetF and pCas plasmids can be subsequently cured from the bacteria. Although developed ... the transcript. Different Cas orthologues and subtypes function somewhat differently and can be more ...Cas9 (dCas9) alone or a dCas9 fused to the omega subunit of RNA polymerase, the lab   successfully repressed...such a purpose (Selle et al., 2020).  Targeting a subset of bacteria in a population Work from the Lu lab...

..., it cannot be used for subcellular analysis such as morphology or subcellular localization studies. For...cell? Or perhaps, you need to analyze a specific subset of cells in a complex population. Have you spent...this technology can be used to isolate a specific subset of cells in a population in a procedure termed ...not, however, permeabilize cells and requires subsequent permeabilization with detergents such as triton-x...instruments have specialized components that can divert a subset of cells. During FACS, the sample stream oscillates...each well allowing users to easily isolate and subsequently expand monoclonal cultures.  Interpreting flow...they are interested in and further analyze that subset of cells. An immunologist, for example, might be...

...follows: Cap Subject Industry H1B Academic H1B National Interest Waiver EB-1A EB-1B Cap subject industry...companies to submit an application on their behalf.  If an individual obtains a cap subject H-1B (last ...numerically limited visas are referred to as “cap” subject H-1Bs. The H-1B allows U.S. employers to employ...numerical quota. Four types of employers are not subject to the H-1B cap: institutions of higher education...national’s proposed work endeavor must have both substantial merit and national importance; (2) The foreign...show that you serve the national interest to a substantially greater degree than the majority of your peers...Addgene Blog Transferable Skills Guide 6 Steps to Submitting a Resume That Gets Seen Tips for Writing a Good...