...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 ...

...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...

...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...

...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...

...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...

...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...

...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...

... 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...

...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...

...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...

...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...

...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...

...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...

...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...