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...looking to use antibody-based applications in their research.  Like all our eBooks, the Antibodies 101 eBook...101 blog posts on an antibody-specific topic, allowing you to easily access relevant information for planning...re very new to antibodies as research tools, you may find “Chapter 1: What is an Antibody” quite useful...to find an antibody that works, we highly recommend “Chapter 3: Finding the Right Antibody for Your Experiment...Experiment.” Whatever your antibody application question, the Antibody 101 eBook is a great place to start...excited to share the very first edition of our Antibodies 101 eBook! The latest addition to our eBook collection...

...non-enveloped viruses, so they are resistant to alcohol-based disinfectants. A 10% bleach solution should ...value greatly. Applying this mantra to AAV selection, there are three important factors related to location...and using AAVs have in common? The three most important items when looking for a house and choosing an...location, location! Just like this common real estate mantra, the choice of serotype (which determines which...the cell type under study. Many AAVs will show significant expression in a variety of cell types, but you...retrograde transport) or towards the synapse (anterograde transport), which is a possible mechanism for...and 9, not only do they undergo retrograde and anterograde transport, but they also have similar transport...

...editor would restore antibiotic resistance. To find such a protein, they created a mutant library of E. coli...improving base editing platforms for plant systems.  Figure 2: Plant base editor platforms and their...components. Adenine base editors for plant We developed plant adenine base editor (ABE) vectors to install... Cytosine base editors for plant There are several CBEs available for plant systems. Target-AID based ... Figure 3: Activity windows of plant base editors. Plant CBE and ABE with expanded PAM compatibility...constructed a number of plant base editors replacing the SpCas9 with the mutant versions of SpCas9 and ...the base editing scope in rice by using Cas9 variants." Plant biotechnology journal (2018). PubMed PMID:...

...Ready-to-use recombinant antibodies at Addgene Addgene's Antibody Guide Addgene's Antibody Plasmid Collection...Addgene blog Antibodies 101: Reading a Flow Plot Antibodies 101: Flow Cytometry Controls Antibodies 101: Validation...single-fluorophore-conjugated antibodies. These methods can include labeling markers with multiple proteins/antibodies, targeting... to antibodies and 'staining' to refer to small molecule dyes. Indirect labeling While antibodies used...fluorophore-conjugated antibody commercially available. In other cases, fluorophore-conjugated antibodies may exist...fluorophore-conjugated secondary antibody is added that binds the conserved, host species-specific (“anti-rabbit”) region...region of the primary antibody (Figure 1b). As above, the advantage of this system is increased flexibility...

... from your antibody panel. They are made up of compensation beads and a conjugated antibody. Compensation...your sample. The isotype antibody will be virtually identical to the antibody for your marker of interest...Introduction to Gating in Flow Cytometry Antibodies 101: Flow Cytometry Antibodies 101: Conventional vs. Spectral...Resources on Addgene.org Addgene's Antibody Guide Browse Addgene's Antibody Collection        ...Depending on the markers you are looking at and the antibodies you are using, you may need to include additional... synthetic beads that can bind to a conjugated antibody and beads that are unable to bind to them: like...The compensation beads can bind to any primary antibody you add. Single color controls are used to help...

...differences include the ferrule implants and patch cables. One huge advantage of using rats is their superiority...but, in today’s post, we’ll discuss some of the important considerations you should think about when developing... preference.   Historically, ICSS was done by implanting an electrode and non-specifically electrically...literature to find all the applications of optogenetics relevant to your field. Ultimately, your question should...stimulation could induce seizures or some other unwanted phenomenon or it could reveal something fascinating...could move your project onto the exciting—and unanticipated—road of discovery. Monitoring behavioral output...interested in studying a brain region that may be important in memory, an appropriate behavioral test might...

.... Other relevant blog posts 3 Challenges in Plant Synthetic Biology Quick, Versatile Plant Transgenesis...As previous blogs have noted, plants are an important foundation for life on Earth. Selective breeding...shaped the plants that we grow and eat, and genetic engineering will continue to improve plant nutrition...inversely proportional to size, and many plant scientists want to transfer large genes, smaller binary ... is functional in a variety of plant species. The kanamycin antibiotic resistance gene is controlled by...PMCID: 4045853.  Find other plant resources at Addgene Check out Addgene’s plant vectors! Find the plasmids...nutrition, yield, and pest resistance. Much of plant genetic engineering revolves around Agrobacterium tumifaciens...

...Viva la (bacterial) resistance!  What are antibiotics? Antibiotics are generally defined as agents that kill...common antibiotics used in labs today are semi-synthetic or fully synthetic compounds. Antibiotics can be...the antibiotics listed in the table below. Note, in this post we'll focus primarily on antibiotics against...how to prepare antibiotic stocks, see Addgene's Reference Page. How else can antibiotics be used in the...scientist to select bacteria that are only resistant to the antibiotic that remains intact. Tips and tricks ...Resistance to antibiotics is a widely used tool in molecular biology, yet scientists rarely stop to think...Additionally, the presence of a plasmid is disadvantageous from the bacterium's perspective – a plasmid-containing...

...this logic, ω-dCas9 variants recognizing multiple PAMs should have a fitness advantage over proteins with...flat out better than SpCas9. I want everyone to test it because I want to know the answer", says Liu. ...work, there are other options. Commonly used Cpf1 variants, with a  5’ TTTV PAM, improve targeting in AT-...respectively. Researchers also engineered SpCas9 variants with alternative PAMs like NGAG and NGCG and SaCas9...phage-assisted evolution to generate and select for Cas9 mutants with PAM flexibility. In this system, the mutagenesis...promoting survival of only the broadest-targeting variants. For the final round of evolution, they used a... a no-G PAM library to enrich for Cas9 variants with activity on non-NGG PAMs, generating xCas9 3.0-3.13...

...Reactions Anti-drug antibodies (ADAs) are formed when the body recognizes a drug as a foreign antigen, triggering...replicate the complexity of antigen processing events that take place within an antigen-presenting cell like ...Beijnen, J. H., & Schellens, J. H. (2016). Antidrug Antibody Formation in Oncology: Clinical Relevance .../s12929-019-0592-z  Resources Antibodies 101: Introduction to Antibodies CRISPR in the Clinic   ...Monoclonal antibody drugs are popular therapeutics for a plethora of disease conditions, from cancer ...cancer to autoimmune disorders. Antibodies administered as drugs are still immunogenic, meaning that they elicit...of immunogenicity can help evaluate risk of an antibody.  In silico prediction of adverse reactions In...

...common antibiotic resistance genes and which applications are best (or worst) for each antibiotic. The... as SHuffle cells, are already resistant to this antibiotic due to stable integration of resistance genes...-in-one antibiotic — it is usable in eukaryotic cell culture, yeast, bacteria, and even plants! The zeocin...host DNA damage. Choosing an antibiotic resistance Choosing the best antibiotic resistance ultimately comes...pay attention to antibiotic selection. If you feel like multiple of the above antibiotics would work for... the pros and cons of each antibiotic and happy selecting!   Antibiotic Resistance Stability Time ...Over-Agar Antibiotic Plating Plasmids 101 eBook Resources on the Addgene blog Plasmids 101: Antibiotic Resistance...

... blog Antibodies 101:Selecting the Right AntibodyAntibodies 101: Secondary AntibodiesAntibodies 101: Buffers...Addgene.org Addgene's Antibody GuideAddgene's collection of ready-to-use recombinant antibodies...a given experiment, it is important to consider how many parameters you want to use in a given experiment...detector using a series of mirrors and filters. Importantly, conventional flow cytometry uses one detector...conventional and spectral flow cytometry, it is important to consider the expression levels of the markers...two highly overlapping cellular markers, it is important to pair them with two fluorophores that have distinct...is critical that your controls and your sample antibodies are from the same batch, to avoid unmixing issues...

...ahead of time that you want to probe your blot with multiple antibodies. You’ll want to set up your blot ...starting with the least-abundant protein and working toward the most abundant. If the abundance of your...isn’t too different, consider antibody affinity: stronger binding antibodies will require harsher stripping...without a re-wetting step. Important: do not dry your membrane with antibodies on it if you plan to reprobe... avoid retaining residual antibodies. If you have already applied antibodies and need to dry the membrane...stripping solution uses low pH to disrupt antibody binding to antigens. Ingredient Concentration or ...you don’t see any signal, you’ll want to check for the primary antibody by re-blocking your membrane and...

...expression of antibodies in plants using syringe agroinfiltration." Recombinant proteins from plants. Humana...transgenic plant can take even longer, a transient plant leaf transformation process could save the plant biologist...Browse our Plant Plasmids and Resources page Find synthetic biology plant plasmids Find CRISPR plant plasmids...vincristine, are regulated in the medicinal plant, the Catharanthus roseus (Figure 1). The ultimate goal of...to enhance their production either in the plant or in plant tissue cultures.   C. roseus is not a model...pathway. Why plant scientists do transient leaf transformations The development of transgenic plants is a common...the plant sciences “transient transformation” usual means that neither a stable cell line or plant is ...

...Addgene blog Antibodies 101: Flow Cytometry  Antibodies 101: Reading a Flow Plot Antibodies 101: Conventional...case through conjugated antibodies targeting the cell surface markers CD11 (anti-mouse CD11b-BV510), which...looser gate here. I am not worried about including irrelevant events in my analysis, as these will get excluded...Gating Having gated my single lymphocyte cells, I now want to exclude all dead cells from my analysis. For ... protect themselves from it. So here, I actually want to gate on my negative, unstained population, which...experimentally-specific gates. Two-dimensional staining (quadrant gating) For our last example plot, I will show...present on dendritic cells but not T cells, and CD8 (anti-mouse CD8b-PerCP-Cy5.5), which is present on cytotoxic...

...bacteria, it is important to note that the term ‘antibiotic’ can also be used to describe anti-parasitic drugs... is not naturally resistant but has become resistant somehow overtime. One fantastic example of intrinsic... spread of antibiotic resistant bacteria, check out this resource on preventing antibiotic resistance ...Addgene's Microbiology Pages What is antibiotic resistance? Antibiotic resistance is defined as microbial... to antibiotics designed to kill them or to impede their growth. When we hear the word ‘antibiotic’, very... drugs and anti-fungal drugs, because ‘antibiotic’ merely means that the drug prevents the growth of a...treat. Learn How Antibiotic Resistance Genes Are Used in the Lab How does antibiotic resistance develop...

...improve your grant writing skills. Good luck! Further advice on grant writing Since successful grant writing...with your work, your advisor may want to mentor you during the grant application process. When my PhD ...have no idea how to write a grant!”, “I don’t have time for this if he wants data by next week!”, and “...RFA contains all of the important information required for a successful grant application. It typically... advice on writing a grant proposal. "Murder Most Foul: How Not to Kill a Grant Application" by Vid Mohan-Ram...How to write a research project grant application" by the NIH Still want more information on the application...their advisor alone applies for these competitive grant applications; however, in some cases, your advisor...

...GreenGate – offers all of those same advantages to researchers working in plant model systems. “The cloning is... most frequently used elements in plant expression cassettes: plant promoters, N-terminal tags, coding...C-terminal tags, plant terminators and resistance cassettes for selection of transgenic plants. The complete... in vitro; it’s no different in mouse, human or plants,” explained Jan Lohmann of Heidelberg University...based on the daily needs of an advanced molecular plant science lab.” Introducing GreenGate As Lohmann ...required to generate multi-construct transgenic plants. In the most practical terms, what GreenGate can...Arabidopsis, it should in principle work in any plant species. Tricks of the trade Lohmann does have one...

...competitive ELISA use antibodies coating the wells to capture antigen from a sample, whereas antigen is bound directly...as a reference antigen — that can also bind to that antibody. Then, the protein-antigen mixture is added...multiple secondary antibodies can bind to a single primary antibody. The secondary antibody is conjugated ...blog Learn about the advantages and disadvantages of using monoclonal antibodies. Read about other techniques...that any antigen within a sample adheres to the well (either directly or by binding to an antibody that is... coated with the antibody. Then, you add your sample to the wells, and the antibodies bind to your protein...is the addition of a detection antibody (typically a monoclonal antibody) that also recognizes the same...

...Fragments. Antibodies 2:193–208 . https://doi.org/10.3390/antib2020193  Khantasup K, Chantima W, Sangma...Comparison between the IgG antibody and scFv.   What are the advantages and disadvantages of a scFv vs. a monoclonal... monoclonal antibody? Advantages of scFvs: scFv’s advantages are rooted in their size. Being smaller means...the ways antibodies are. scFvs are even used to screen for new antibodies. To do this, antibody variable...vector encoding the antibody constant domain scaffold to convert the scFv into an antibody.  The ability to... Prospects of Antibody Fragments. Antibodies 8:28 . https://doi.org/10.3390/antib8020028  Bird R, Hardman...When you think of antibodies, you probably think of monoclonal antibodies (mAbs). mAbs have been around...