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...blot: we meet again. So useful, yet so finicky to design and optimize. Today we’ll cover the normalization...normalization can amplify other errors in experimental design and gel quantification. Figure 3: In a perfect...final blot. Quantifying the entire lane of protein provides a loading control that can be used for normalization...Antibodies 101: The Basics of Western Blotting Technical Design of a Western Blot Troubleshooting and Optimizing...

...Applying for a Faculty Position: the View from Both Sides," are summarized below. Check Out Erik Snapp's ...You want people to focus on you and your ideas. Slides will be     distracting. What is the big question...feedback. Many of these documents are too vaguely described in other sources of guidance or the document examples...single malt whiskies, but I remember the days of modest postdoc salaries all too well. If there's a group...

...cofactor binding proteins as described above. The smallest tags are peptides that are engineered to bind...homology domains can be used to detect specific phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production...Péter, and Tamas Balla. "Live cell imaging of phosphoinositides with expressed inositide binding protein domains...

...expression to specific tissues and cell types. If desired, spatiotemporal control of transgene expression...systemic AAVs to find commonly used vector cargo designs and administration doses. Note: If capsid immunogenicity... Table 1 gives a brief overview of published descriptions of commonly used or emerging capsids. Selected..., B.E., Gradinaru V. Systemic AAV vectors for widespread and targeted gene delivery in rodents. Nature...Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain. Nature Biotechnology... prioritization for cell type-specific enhancer design. eLife,11, e69571. https://doi.org/10.7554/eLife...

...every item I tested was constantly tracked with barcodes and spreadsheets. It was great to be in the lab...industry positions. There were, however, some downsides to working in a large commercial company. In my...aspects of the company and gave me the freedom to design new projects and grow in the role. I understand... job different every day. I love the freedom to design and develop my own projects, but working remotely...

...brain. The cause of this neuron death is unknown. Despite the idiopathic (having no specific known cause)...scientific community at low cost, with rapid delivery. Includes antibodies, viral vectors, animal models, purified...

...response element (RRE), an internal promoter, and the desired gene (transfer vector plasmid), 2) the HIV-1 genome...process is to test several protocols that have been described (protocol 1, protocol 2, protocol 3) to find the...quantify viral RNA. All of these methods have been described in a previous Addgene blog Once you know your ...Miller, A. DUSTY, and C. A. R. O. L. Buttimore. "Redesign of retrovirus packaging cell lines to avoid recombination...

...uncertainty of it all. Though I had spent the first few decades of my life following a very straightforward conveyor... harassment in academia or a large company who decides how they are going pursue regulatory approval for...their Local Engagement Playbook for step-by-step guides on how to engage in your local community.  Read...comprehensive resource developed by Dr. Kelly Singel which provides an overview of all available science policy fellowships...

...to be present and recognizable by your antibody despite the manipulation. Additionally, not all sample ...large deletion that removes the entire exon that codes for the epitope recognized by your antibody. You...specific, then you expect that the bulk of the peptides identified to be from the target protein. This...tuned for ready-to-use antibodies from Addgene Designing a recombinant validation experiment? Find plasmids...

...end-joining, but will also efficiently ligate many undesirable structures, including substrates containing gaps...fidelity") better than T4 DNA Ligase (21, 25, 26). Despite this higher fidelity, Taq DNA Ligase can still ...possible NN pairs in the template. The probes were designed such that each possible pairing resulted in a ... high fidelity ligase, along with careful probe design and reaction condition optimization, will be critical...

...above 600 nm. LumiFluors Schaub, F.X., et al. (3) designed two imaging probes. One consists of NLuc fused...Reporters. The LumiFluor reporter constructs were designed to be packaged into a retrovirus for infection...BRET probe for imaging. These authors set out to design a better fluorescent protein that could be co-imaged...acceptor, the more powerful the FRET signal. In well-designed FRET-based biosensors, the intensity of the FRET...

...update, we have added new sections to share and describe some incredible new CRISPR technologies developed...much-anticipated prime editing section and additions describing CRISPR’s ability to make large genome edits using...transposases or the Cascade-Cas3 system. We also describe newly engineered Cas9s with more flexible requirements...

...When performed correctly, fluorescence microscopy provides a high contrast image in which a bright signal...specificity to the viewer. Finally, modern microscope designs may further exploit the unique characteristics ...one object in the final image. Over the past two decades a number of techniques have been developed that...resolution and decreased phototoxicity. Lightsheet designs that allow for multi-view imaging can image thick...

...been infected with a microbe, to trigger immune cascades that clear these infections. Anything that generates...immune response and initiates different immune cascades. In the research setting, antibodies of different...antibody, which recognizes the primary antibody and provides the method of detection. You’ll also want to use...used for. Addgene’s Antibody Plasmid Collection includes plasmids that express antibodies, nanobodies, ...

...eBooks, the Antibodies 101 eBook is designed to be a helpful desktop reference. Each chapter brings together...an Antibody” quite useful. If you’re looking to design your first flow cytometry experiment, “Chapter ...

...occurring earlier but that the photo evidence was destroyed to protect the innocent revelers. Our Halloween...for Halloween and how seriously we take costume design. In case you need more convincing, I’ve got just...to great customer support by dressing up as our Zendesk email system.  Our 2013 winners dressed up as ...

...single gene KO and pooled screening approaches; (4) encodes a Thy1.1 selectable marker to facilitate the isolation...assess T cell responses and clonal dynamics in lymph nodes (LN) and tumor cells. Figure used with permission...also reveal novel pathways. The FITS deposit also includes pLX_305_Ovalbumin, a plasmid that enables introduction...

...and the Addgene Help Center. A big part of this redesign was pulling all of our science options into a ...Addgene’s website easier to navigate, thanks to design choices informed by Web Content Accessibility Guidelines...word for it — check out all the new features and designs addgene.org! And when you’re finished, feel free...

...2022, from the launch of a new material to the redesign of a community resource. Now that we’re in the...antibodies. Everything in this collection, which includes both neuroscience and general-use antibodies, ...website saw 2.2 million visitors from 231 countries. Guides and protocols were our most popular content, while...

...Feng Zhang’s lab at the Broad Institute and was designed and cloned by Neville Sanjana. This fantastic ...your gRNA of interest to mammalian cells. It was designed to have increased lentiviral titer as compared...sampling of the research it has already been used for includes work probing mTOR signaling in cancer cells, deciphering...