We narrowed to 897 results for: ANT

...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...

...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...

...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...

...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...

...these enzymes implies that even a minute quantity can significantly reduce the operational time for a given...efficiency.  Finding recombinant psychrophilic enzymes for the host The expression of recombinant proteins for... contained cold tolerant chaperones (the Cpn 10/60 isolated from Oleispira antarctica, and the P. haloplanktis... psychrophilic proteins. The primary problem we wanted to overcome was inefficient protein folding at ...well under suboptimal conditions. This means recombinant proteins can aggregate or precipitate out as ...would allow the stable folding and expression of recombinant psychrophilic proteins. As the figure below shows.... And thus was born ELSA - our happy lil’ cold-tolerant E. coli! Additionally, we’re also working towards...

...about and jumping into the discussion! Semantic Scholar Semantic Scholar is an AI-backed literature search...Intelligence. Semantic Scholar analyzes publications and figures using machine learning to extract important features...return results that are more relevant to your search terms. Key features Semantic Scholar has several ways...millions of them. The real challenge is finding relevant papers. The latest installment of the Early Career...to find papers  You’ll need to start following relevant accounts in order to turn Twitter into a paper-finding-machine...keywords. It’s sort of like a keyword search that’s constantly updated. This Twitter list contains >300 Twitter...at the top of this filter. A unique feature of Semantic Scholar search results are the topic pages, which...

...controls needed for relative quantification of a western blot — and why you might want to be careful relying ... Loading controls are even more important for semi-quantitative western blots. No pipetter is perfect...blotting quantification” (Gilda & Gomes, 2013). To address this source of error, it’s important to determine... on so called “housekeeping proteins.” Fully quantifying a western blot requires a standard curve with...cases. Often, you may not need to know the exact quantity of your POI, but simply whether its concentration...experimental parameters. This is a job for a semi-quantitative western blot. Most western blots benefit from...accurately compare different lanes? In relative quantification, the POI bands are normalized to the loading...

...step where the target antigen binds to antigen-specific antibodies. The antibodies are conjugated with ... then adds an antigen-specific detection antibody.  Importantly, the detection antibody and the capture...Addgene blog Antibodies 101: ELISAsAntibodies 101: Selecting the Right Antibody Antibodies 101: Polyclonal...Polyclonal Antibodies Resources on addgene.org The Addgene Antibody GuideAddgene's recombinant antibody collection...primary antibody first binds to the antigen and then a reporter enzyme conjugated secondary antibody binds... to the antigen in a sample and then a second detection antibody binds. The detection antibody can be ...The more antigen in the sample, the less antibody available to bind to the competing antigen. In direct...

...use two antibodies - one against your target of interest and a second against the first antibody. When ...same species as your primary antibody are present, your secondary antibody recognizes both the endogenous...endogenous IgGs and your primary antibody, resulting in significant background staining (Figure 1A). Not all...because the secondary antibody is not able to distinguish between the primary antibody and endogenous IgGs...secondary antibody bind endogenous IgGs? Don’t use one. If it is possible, using a primary antibody that is...and secondary antibody complexes (Figure 2B) Some labs have found that you can significantly decrease background...samples as homogeneously as a single antibody. Using a secondary antibody that is a Fab fragment, single-chain...

... What are toxin-antitoxin systems? These toxic substances are part of toxin-antitoxin systems that are... while the antitoxin is unstable, meaning that the cells must continually produce antitoxin to avoid cell...will still inherit some toxin and antitoxin. However, the antitoxin will be degraded leaving the cell ...applications. The discovery of toxin-antitoxin systems Toxin-antitoxin systems were first discovered in 1983.... The encoded toxin-antitoxin system consists of CcdB (toxin) and CcdA (antitoxin) where CcdB inferes ...higher percentage of transformants contain the ccdB-less construct. Other toxin-antitoxin systems and their...-based toxin-antitoxin systems. One of the most well known protein-based toxin-antitoxin system is MazE...

...fluorophore-conjugated antibodies that attach to a specific target. By using a mix of different antibody-fluorophore...a conjugated antibody to detect CD45, and I'll need to choose the fluorophore my antibody is conjugated...the Addgene blog Antibodies 101: Introduction to Gating in Flow Cytometry Antibodies 101: Flow Cytometry...Cytometry Antibodies 101: Flow Compensation Resources on Addgene.org Addgene's Antibody Guide Ready-to-use ...Ready-to-use antibodies in the Addgene repository Antibody Protocols ...distinguishing live from dead cells. The combination of antibody-fluorophores, fluorescent reporters, and/or dye...excitation and emission wavelengths, it is also important to know the brightness of individual fluorophores...

...third Addgene/Seeding Labs plasmid grant and plans to use the grant to order plasmids she can use as positive...the Addgene/Seeding Labs Plasmid Grant to a third round of applicants at the start of 2017 and would like...in PCR-based cervical cancer screens. This third grant represents the continuation of our efforts to provide...Bengyella and Kwabena Duedu who were awarded similar grants back in June, Gladys will be using plasmids in ...caused by persistent infection of HR HPV types, predominantly HPV 16 and 18.” HPV plasmids as positive controls...Superhero 5k Find New Cancer Resources Learn About Our Grants to Rare Disease Research Resources on Addgene.... seeding labs plasmid grant...

...requires the presence of dsRNA. In plants, CRISPR technologies are predominantly used to disrupt function at ...from an easy method to identify and quantify actively silencing plants. Non-destructive methods, most popularly...silencing plants can be easily identified and have readily observable phenotypes quantified. Fluorescent...designed and widely implemented in plants, such as in the model flowering plant Arabidopsis thaliana (Li et ...possible in plants outside of limited, single cell protoplast analysis. Instead, microscopic plants must be...consuming, as RNAi mutants with a reduced growth phenotype will require additional plants to have enough ...MicroRNA Screens for Effective Gene Silencing in Plants. Plant Cell 25:1507–1522 . https://doi.org/10.1105/...

....  In conclusion, it is important for you to establish in advance how you want to utilize your plasmid...purifying a plasmid from a bacterial culture, it is important to consider your experiment.  It will dictate ... a set of techniques that are used to insert recombinant DNA from prokaryotes or eukaryotes into a synthetic...approach you choose ultimately will influence the quantity of DNA that is required and, as a consequence,...To allow efficient and flawless cloning it is important that the preparation is pure from other nucleic...(genomic DNA and RNA), proteins and chemical contaminants that have been used during the purification ...scientists an indication of purity versus protein contaminants. The 260 and 230 nm ratios should fall ~2 - ...

...some plant-specific changes to CRISPR components are necessary to enable genome editing in plant cells... allow for the use of CRISPR in plants, and outline the various plant genome engineering tools that are...system in plant cells. Like other model systems, expression of S. pyogenes Cas9 or Cas9 variants (hereafter...question. In plant cells, gRNA expression is achieved by placing the gRNA downstream of a plant-specific ...optimized Cas9 variants have been created in an effort to increase translation in a particular plant species ...target genes in plant cells, respectively. Cas9 expression is commonly driven by plant-derived RNA pol... editors have been developed for plants and prime editing for plants has been developed for rice by Yiping...

...Other labs have used PCR-based screening to detect mutants, but this method limits screening to a few hundred...

...a monoclonal antibody against a target antigen. This contrasts with polyclonal antibodies, which are mixtures...a repository for monoclonal antibodies, distributing over 3,000 antibodies and associated hybridoma cell...distributing monoclonal antibodies at cost. We distribute over 65,000 antibody units per year from our...monoclonal antibodies, which are each produced by a single B cell. To create a monoclonal antibody cell line... of monoclonal antibodies obtained from serum.  Our most popular monoclonal antibodies include those ...Monoclonal Antibody Research Institute dedicated to: 1) developing new ways of generating antibodies, for example...through the use of less toxic adjuvants; 2) generating complex antibody chips that allow assessment of...

... genomes so as to make plants, and agricultural crops in particular, resistant to disease. They also devise...them. The MoClo Plant Kit makes it really easy to clone genes for expression in plant cells. Sylvester... lot. The problem in plant science is there is a paucity of resources. We wanted to expand the toolkit...also gives quantitative data for regulatory elements, which has not been available for plants. Addgene:...tools for editing plant genomes. We will definitely plan to share those tools with the plant community. We...Remember plants. Every time you look at your plate, think about how much that’s on it are plants, and the...animals you eat ate plants too. Think about how many people are starving. I think plant biology is massively...

...gene of interest and keeping its expression level constant. In the case of the stabilized promoters from ...expressed. What’s the use of stable gene expression? Constant gene expression can enable more confidence in ...Christopher A. Voigt. "Engineered promoters enable constant gene expression at any copy number in bacteria...

... address the growing clinical problem of antibiotic resistant bacteria. See the picture to the left for... drug resistantance genes in the bacteria thus making the bacteria sensitive to known antibotics. These... able to participate when the organizers of the Canton Nucleic Acids Forum (CNAF) 2015 conference invited...scientists still working at the bench. It was a fantastic opportunity to talk to so many scientists who ... of plasmid requests from Addgene. Addgene - Canton Nucleic Acids Forum 2015 from Joanne Kamens, PhD...mammals, expressed in neurons, and are especially abundant in neuronal synapses. shRNA silencing of one circular...his list of the bacterial species that “ESKAPE” antibiotic treatment resulting in major problems in hospitals...

...Jennifer Tsang. CRISPR-Cas technology is constantly evolving. Variants of Cas proteins can be used for genome...proteins. Discovery of anti-CRISPR proteins That is, until the discovery of anti-CRISPR proteins in 2012...DNA. Since these early anti-CRISPR studies, scientists have discovered over 50 anti-CRISPR proteins that...they block can be found here. How do anti-CRISPR proteins work? Anti-CRISPRs proteins are highly diverse...2012 (Bondy-Denomy et al., 2012). Anti-CRISPR (Acr) proteins are phage-derived small protein inhibitors...the Sontheimer and Davidson labs discovered three anti-CRISPRs for a type II system in Neisseria meningitidis...genome editing. In this study, the labs found that anti-CRISPR prevents Cas9 activity in HEK293 by reducing...