...their recombinant antibodies. Since recombinant antibodies are a newer technology, it is important to show... blog Antibodies 101: ValidationSelecting the Right AntibodyPlasmid-based Recombinant Antibodies    ...and cost-effective) antibody for my experiment – the “goldilocks antibody.” Antibodies are one of the most... the antibody crisis, one must corral and corner the goldilocks antibodies. A goldilocks antibody is highly... popular and useful antibodies are, they are in a midst of a crisis. The “antibody crisis” came to light...light as antibodies became commercialized, revealing the unreliability of antibodies. There are three main...ability – i.e., the antibody can no longer bind to the target). Unreliable antibodies cause research that...

... developed Nano-lantern technology. Nano-lanterns contain a Renilla luciferase variant fused to an FP;...contexts. Improving Nano-lanterns In 2015, Takai et al. expanded the Nano-lantern color palette to include...producing a constant supply of substrate. Using this compound to excite the Nano-lanterns, Takai et al... Nano-lanterns (eNL) described in Suzuki et al. improve the brightness of previous Nano-lanterns by 2-...appropriate for use with Nano-lantern technology. Multicolor imaging using Nano-lanterns requires a linear unmixing... signals of various Nano-lanterns, but it does not isolate each Nano-lantern, so Takai et al. used a linear...explained further in Takai et al. Nano-lanterns and ready-made Nano-lantern biosensors are available from Addgene...

...library to a significant degree using different types of AAV. Expression Another advantage of AAV vectors...ones. In those cases, AAV vector screens may be AAVantageous!       In vivo, in vitro, ex vivo CRISPR screens...cells with the CRISPR library in vitro before transplanting them into mice. For in vivo screens, mice are...locations that could cause deleterious effects. It is important to note, however, that while AAV rarely integrates...the amount of cloning, another option is to use variants of Cas9, like split Cas9, or a smaller Cas9 ortholog...Laboratory). https://doi.org/10.1101/2023.06.13.544831 Santinha, A. J., Klingler, E., Kuhn, M., Farouni, R., Lagler...

...animation is here to help! "Antibodies 101: Antibodies in the Lab" shows how antibodies are used to study proteins... Addgene Addgene's Antibody Guide What is an Antibody? animation Addgene's Antibody 101 eBook    ...Thinking about using antibody-based applications in your experiment...but not sure where to start or ...introduction to — or quick refresher on — these common antibody applications.  Thanks for watching, and we hope...

...which antibody to request for your experiment!  Addgene’s Antibody Data Hub Addgene’s Antibody Data Hub...Updated AAV Data Hub Antibodies 101: Validation Antibodies 101: Selecting the Right Antibody...the Addgene antibody collection can look at user-deposited data detailing how an antibody performed in...Figure 2: Search options in the Antibody Data Hub. Data reports  Each antibody submission is used to create...immunocytochemistry experiment performed with the anti-Lhx6.1 antibody.    For a more detailed understanding of... the antibody performed.  Figure 5: Results for the data report shown in Figure 3. The antibody is rated...requesting an Addgene antibody, the Data Hub can help you decide if an antibody is a good candidate for...

...reagents, such as recombinant or hybridoma-derived monoclonal antibodies. It is also important to seek out ...Synaptotagmin-1 antibodies and corresponding Multi-rAb CoraLite® Plus 647 secondary antibodies. Antibody staining...0.35 µg/mL (1/100). * = monoclonal antibody, ** = recombinant antibody. Figure adapted from Biddle et al...multiple antibody clones targeting the same protein (independent antibodies). If two antibodies, that recognize...Good Antibodies, with significant input from Michael Biddle, PhD, also from Only Good Antibodies. The ...2023). Scaling of an antibody validation procedure enables quantification of antibody performance in major...Antibody validation is to confirm (or refute) that the antibody is selectively detecting the target-of-interest...

...regions of the antibody that actually bind antigens.      Figure 1: IgG antibody structure with...portion of the antibody. Fab fragments are useful when you just need an antibody’s antigen binding function...   Why Fabs?  Antibody fragments have a handful of advantages over full antibodies. Their smaller size...as primary antibodies, you can make your own antibody fragments as needed.  In contrast, anti-IgG antibodies...secondary antibody recognizes endogenous antigens. In these cases, even though one full antibody may be ...Prospects of Antibody Fragments. Antibodies 8(2):28. https://doi.org/10.3390/antib8020028    Wessel GM...However, sometimes you want a tool that is a little more compact — more of an antibody fragment rather than...

...your samples with multiple antibodies, the fluorescent probes on the antibodies may have similar emission...Introduction to Gating in Flow Cytometry Antibodies 101: Flow Cytometry Antibodies 101: Conventional vs. Spectral...this handy emission spectra chart to find your antibody's spectra). Your cytometer will therefore record...BioLegend’s Fluorophore Guide Chart.  Pro tip! Some antibodies go by different names depending on your flow ...

...heavy-chain only antibodies in the serum of a camel and soon after found similar antibodies in llamas and...Comparison of a heavy-chain only antibody and single-domain antibody, under the commercial "Nanobody" ...to other antibodies and affinity reagents. But for a quick refresher, some of the advantages of sdAbs ...conventional primary antibodies, and there are unique benefits to using sdAbs as secondary antibodies. sdAbs, however...single-domain antibodies genetically fused to fluorescent proteins and used to visualize target antigens in living...single-domain antibodies without the need for a species that naturally produces heavy-chain only antibodies. (A)...heavy chain only antibodies) (B) Nanomice express hybrid heavy-chain only antibodies that are the source...

...resources on the Addgene blog Antibodies 101: Introduction to Antibodies New Antibody Animation ...bite-sized portions of antibody concepts. Each short video answers an Antibodies 101 question, like "what...what is an antibody?" or "what is an IgE isotype?"  @addgene Ready to learn about antibodies? Our new ...new antibodies animation has all the information! Link in bio. #STEM #antibodies #molecularbiology #science...Our popular Antibodies 101 animation is becoming a TikTok series! Follow @addgene on TikTok for bite-...

....org  Antibody GuideRecombinant antibodies available from AddgeneBrowse our recombinant antibody plasmid...as is antibody recognition, since antibody-specific antibodies bind to and recognize the constant region... (the antibody region that binds to the antigen) from existing antibodies. Each cleaved antibody will ...from camelid antibodies to antibody fragments to tools completely unrelated to antibodies.  Why use an...co-crystalize, a major advantage over antibodies (Kwon, 2019).  Single-domain antibodies (sdAbs), sold under...colors.  To antibody or not to antibody?  That is the question… or is it? This post isn’t meant to convert...101: Aptamer FluorophoresAntibodies 101: scFVs Antibodies 101: Introduction to Antibodies Additional resources...

.... Name Our New Antibody Friend We need your help naming our latest mascot, an antibody joining us on ...following along on our journey to name our new antibody mascot, who's joining us in preparation for our...for Abi as they help us prepare for our upcoming antibody launch in late March!      *If you submitted ...Blugene and Aavery's newest friend, our Addgene antibody mascot, here to help us with our upcoming affinity...suggestions here or by using the hashtag #AntibodyMakesThree on Twitter. As with Blugene and Aavery, we...

...s blog Antibodies 101: Monoclonal Antibodies Antibodies 101: Polyclonal Antibodies Antibodies 101: Introduction...Introduction to Antibodies Resources on Addgene.org Find plasmids encoding recombinant antibodies at Addgene...collection of antibodies in your lab. Perhaps you’re even in that mythical place where antibodies are well-...hides the epitope that the antibodies used to recognize the protein, your antibody will no longer be effective...immunohistochemistry assay, you may want to rule out monoclonal antibodies that have been validated solely...Validation  Once you’ve selected an antibody, or multiple antibodies, you’ll need to validate them for ...which antibodies are suitable for which assays and conditions when you’re building your antibody library...

...single antibody to visualize their target. These antibodies are generated against a specific antigen and ...two antibodies: a primary and secondary. For indirect IF, the primary antibody targets the antigen of ...each antibody performs differently and some antibodies impact the performance of other antibodies. But...secondary antibodies on hand to mix and match with a larger collection of primary antibodies. This strategy...target and sample type. Primary Antibody This is the first antibody applied to your sample, which will...incubation time. Secondary Antibody Unsurprisingly, this is the second antibody you apply, which binds ...experimental context.  Choosing antibodies for your assay The antibodies you choose for your experiment...

...expression, measured using antibody conjugated fluorophores Fluorescent antibodies get around the challenges...staining a primary antibody binds to a target and a fluorochrome-conjugated secondary antibody binds to the ...are low as multiple secondary antibodies can bind to a primary antibody thus amplifying the signal.   ...should also include a secondary antibody control in which the primary antibody has been omitted from the staining... about antibodies Resources on Addgene.org Visit the Fluorescent Protein Guide Find antibody plasmids...Have you ever wanted to measure expression of your protein of interest in a single cell? Or perhaps, ...surface receptor are stained with a fluorescent antibody against the receptor and analyzed on a flow cytometer...

...limited understanding of plant biology is a significant technical obstacle to plant synthetic biology. We...pigments (anthocyanins), and the plant circadian clock. Petunias have been a model organism for plant research...here. The first transgenic plant was engineered over 30 years ago, but plant synthetic biology is still...because working with plants can be challenging, there are a lot of unexplored areas in plant biotechnology that...themselves if they want this beautiful flower in their front yard or not. By developing plant aesthetics with...pivot towards plant biotechnology and provide a much needed boost to basic molecular plant research, more...the lack of resources associated with plant synthetic biology. Plant molecular biology has traditionally...

... to embryonic stem cell use and has the major advantage that iPSCs can potentially be derived from patient-specific...mutation and minimize tumorigenicity, an inherent disadvantage of iPS cell technology. Browse Addgene's collection...create cell lines with endogenously-tagged gene variants or browse Addgene’s entire collection of Stem ...and Xeno-Free Culture Conditions: A Clinically Compliant Approach. Stem Cells Transl Med. 2015 Mar 5. pii...Mouse Direct reprogramming of fibroblasts into antigen-presenting dendritic cells. Sci Immunol. 2018 Dec...Lentiviral Mouse Direct Induction of the Three Pre-implantation Blastocyst Cell Types from Fibroblasts. Cell...

...both dividing and non-dividing cells without a significant immune response. These viruses also integrate...that encodes genetic material that the researcher wants delivered to specific target cells. This genome ... See article for more cre-lox shRNA expression variants. Jacks 19319 pLJM1-EGFP 3rd CMV-driven EGFP fusion...destination plasmid for cDNA expression Ramalho-Santos 24129 pULTRA 3rd bi-cistronic expression of EGFP...fusion. See plasmids 36083 and 36086 for other FP variants. Tsoulfas 41393 pCW57.1 3rd Inducible lentiviral...

...science stories Zombie ants Ophiocordyceps unilateralis is a fungus that infects ants and controls their ...atrophy. After the ant’s death, the fungal fruiting bodies being to sprout out of the ant’s body. The fruiting...their behavior. These ants become “zombies” and leave their foraging trail. They climb upwards until they ...temperature and humidity favors fungal growth. The ants bites down and locks their jaws into a leaf, and... onto the forest below for the next unsuspecting ant to stumble upon. Vampire bacteria Vampirovibrio chlorellavorus...genes were appropriately named as spook, spookier, phantom, disembodied, shadow, and shade. Tongue-eating ...

...Addgene The Antibody Guide Antibodies 101: Isotypes Antibodies 101: Choosing the Right Antibody  ...polyclonal antibody is better for your experiment. Monoclonal antibodies contain a single antibody clone and...anti-mouse antibody. Others recognize specific isotopes within that species, like an anti-mouse IgG antibody. Some...subclass specific, like an anti-mouse IgG2a antibody. When using a monoclonal antibody be sure to use a highly...general anti-mouse IgG. In a broadly specific secondary antibody only a fraction of the antibody molecules...example, some goat anti-mouse secondary antibodies recognize some rat primary antibodies (Erickson 1993)....primary antibodies raised in different, and distantly related, species, and secondary antibodies raised...