...different species, or at least from different IgG subclasses, so that secondary antibodies exclusively recognize...Must use different species and/or different IgG subclasses of 1° for multi-color staining Can be expensive...limited cross-reactivity between different IgG subclasses and different species, nanobodies are useful ...require using primary antibodies from different IgG subclasses. Instead, you can pre-incubate individual primary...antibody: species it was raised in and its IgG subclass. Once you know the answer to these two questions...right nanobody for your work. Species Antibody Subclass Nanobody Addgene Plasmid Rabbit N/A* TP897... (3x cysteines) *Rabbits have only one IgG subclass.   Express the Nanobody In Bacteria: Table 2 also...

...encourage you to submit data showing how the antibody worked in your application. The submission process is ...Antibody Data Hub. Data reports  Each antibody submission is used to create a data report, which contains...data showing how the antibody performed in the submitted application, along with a brief description of...reports Just like our AAV Data Hub, each data submission we receive is checked by Addgene's curation team...so you can work through it at your own pace. To submit, simply click "Contribute Data."    Figure 6: ...

...incremental, ordered cell lysis. E. coli cells are subject to lysis by freeze-thaw if not suspended in cryoprotectant...

...against the cellular ribonuclease P/MRP 30 kDa subunit (RPP30) are used as a control for normalization...

...s do it!'" Working as a subject matter expert Caitlin started out as a subject matter expert (SME) at ...more about Caitlin Runne-Janczy and her job as a Subject Matter Expert/ Product Development Manager at eScience...is often like being a jack-of-all-trades. “As a subject matter expert in the educational sphere, it can...it is researching the different aspects of the subject you're writing and then condensing that and making...Hutchinson Cancer Research Center. And make sure to subscribe to the Career Channel of the blog so you don’t...

...if not for this specific submission, then for the submission to the subsequent journal.  As a postdoctoral...Saudi Arabia. From the perspective of an author submitting a paper, the peer-review seems like another dragon...tolerance, I have been peer-reviewing journal submissions since 2015 for a wide variety of journals. Since...routine that works well for me to evaluate the submitted paper in addition to the journal-specific guidelines...

...serotype did you use and what animal subject did you use? For animal subjects it is helpful to report the genetic...location, titer dose (Example: 1E13 GC/mL) and subject age at time of injection. How long post-injection.... Image: The final and required step requires submission of an experimental image result that you generated...this provides a clear overview of what you need to submit data. Please note that the data hub is currently...viewing or data deposition experience. If after submitting or viewing the data you have any feedback at ...

...introduced into the B. subtilis genome. Mobile-CRISPRi is then transferred from B. subtilis to the bacterium...CRISPRi into a single bacterial strain, such as a B. subtilis, or a narrow range of bacterial species, such ...integrative and conjugative elements in Bacillus subtilis (ICEBs1) system. ICEBs1 carries the genes needed...bi-parental mating. Following excision from the B. subtilis genome, the Mobile-CRISPRi ICE element forms a...integrative and conjugative element in Bacillus subtilis." Plasmid 86 (2016): 14-25. PubMed PMID: 27381852...

...please!)... Abi! Thank you so much to everyone who submitted names*, voted, and helped us on our arduous quest... antibody launch in late March!      *If you submitted the names Abi, Lin, or Blottie (our top 3 choices...joining us on our scientific adventures.  You can submit your name suggestions here or by using the hashtag...and it can’t hurt if it’s also a great pun.) Submissions are open until Feb. 3, 2022.  The Addgenies will...choose the new name! Additionally, anyone whose submission is included in the final four will receive some...

...heat block Approach: Viral RNA is extracted and subject to isothermal amplification by primers that target...cleaves the supplied fluorescently labelled ssDNA substrates. Upon cleavage, the fluorescence is no longer...bath Approach: RNA is extracted from a sample and subject to isothermal recombinase polymerase amplification...-2 in the samples. First, RNA is extracted and subject to reverse transcription. This is then amplified...Polymerase and then transcribed to provide RNA substrates for Cas13. Cas13, upon finding complementarity...between the gRNA and a given fluorescently labelled substrate, gets activated and performs non-specific collateral...

...possible to build gene regulatory RSMs (GRSMs, the subset of RSMs that regulate gene expression) that uniquely...attB-attP pairs on a register to record all permuted substrings of a set of chemicals. Specific examples of arrangements...arrangements that should be able to record all permuted substrings of up to seven chemical inputs are given in Table...population to correctly record each possible permuted substring of the inputs, and that five hour inductions were...cells to correctly record each possible permuted substring of inputs (see Figure S9 of our paper (1)). We...described in the literature (5, 6) and may be substituted for one another in an RSM. The predictability...

...from scientists’ submissions, either their deposited gRNA plasmids or from submission of a spreadsheet... the validated gRNA sequences alone. Sequence submission is welcomed for validated gRNAs that are not ...them to this list. Access the spreadsheet for submitting your gRNA sequences here. Features of the validated...develop and confirm new gRNAs, please consider submitting their sequences (and plasmids!) so that this ...

...activities. Working with proteins requires a substantial amount of properly folded, relatively pure protein...doing some research, notably determining the subcellular localization and the number of disulfide bonds...Tags. Beyond E. coli: purifying from Bacillus subtilis and Pichia pastoris pCri also includes plasmids...than E. coli, specifically the bacterium Bacillus subtilis and the yeast Pichia pastoris. The improved secretory.... pCri vector pCri-13a, designed for use in B. subtilis, contains the MISTIC (membrane-integrating sequence...

...updated! Subscribe to the Addgene newsletter for updates on all things Addgene; or subscribe to the Addgene...available for Packaged on Request service. A small subset of AAV plasmids will have technical or legal restrictions...of choice, pick what size prep they’d like, and submit the request. Request details can be accessed within... 100 µL (2.5 mL), or 50x 100 µL (5 mL).  Once submitted, the request will be reviewed by Addgene for legal...

...with a paired t-test, because they eliminate subject-to-subject variation (the test assesses the sum of differences...an unpaired t-test, which does account for subject-to-subject variation in its analysis (the test assesses...unpaired t-tests Samples that come from the same subject or population - before and after a treatment, for...between pairs). Samples arising from different subjects or populations (a treatment group vs. a control...

...regions. The resulting viral-infected neurons are subsequently photo-stimulated through fiberoptic cannulas...ability to specifically infect defined neuronal subpopulations, and this represents a major downside in optogenetics...electroencephalography evidences of arousal in mice previously subjected to steady-state general anesthesia (Taylor et ...ago, however, it was completely unclear which subpopulation of DA neurons could activate appetitive rather...recorded the electrical impulses of discrete subpopulations of this brain area, demonstrating that it is...and Jamie Maguire. "Characterization of a novel subtype of hippocampal interneurons that express corticotropin‐releasing...

...Kanamycin aminoglycoside Binds 30S ribosomal subunit; causes mis-translation Bactericidal 50-100 ug/...Spectinomycin aminoglycoside Binds 30S ribosomal subunit; interrupts protein synthesis Bactericidal 7.5... Erythromycin macrolide Blocks 50S ribosomal subunit; inhibits aminoacyl translocation Bacteriostatic... Tetracycline tetracyclin Binds 30S ribosomal subunit; inhibits protein synthesis (elongation step) ... ug/mL Chloramphenicol   Binds 50S ribosomal subunit; inhibits peptidyl translocation Bacteriostatic...

...database design (schema).  Registered users can submit new fluorescent proteins, add data to existing ...database is free from the risk of inaccurate data submissions, so FPbase includes a versioning system that ...and to view previous versions.  Currently, all submissions also require a DOI referencing primary literature...published literature as soon as possible after submission. Using FPbase The most obvious use case for FPbase...experience in Python, fork the project on github and submit a pull request. Many thanks to our guest blogger...

... own subclone, they were able to develop a 12-plex multiplex imaging system. Mixing the subclones within...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ... a fluorescent protein coupled to a specific subcellular localization peptide. By combining three different...their work with the “signalome,” a 12-plex mixed-subclone system using mStrawberry, localization peptides...

...Monitor neo-substrates Small molecule degraders may recruit CRBN to nonstandard or “neo-substrates.” We advise...evaluation of degradation of known cereblon neo-substrates (IKZF1, IKZF3, GSPT1, ZFP91, and CK1a). While...While expression of these cereblon neo-substrates varies in different contexts (for example, IZKF1 and IZKF3... to date, we have not observed off-target neo-substrate degradation with dTAG-13. Example experiments ...