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.... 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...

...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-...

...Resources on Addgene.org Addgene antibodies Addgene’s antibody guide Antibody protocols  Resources on the...especially if downstream antibody detection is desired… and yes, we have an antibody for that!   Fig....of these tags also have commercially available antibodies, resins, beads, and other purification aids readily...are trying to purify The expression system If you want the tag to remain on your protein for downstream... to remove the cleaved tag from solution. MBP antibodies are available for detection and/or purification...purification of MBP-tagged proteins. The antibody is an alternative to amylose resin and also provides a visual...you are having solubility issues!). Need a GST antibody? Addgene has you covered! His His tags, often ...

...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...

...Lab, using recombinant antibodies with different isotypes. (A) Neocortex labeled with anti-pan-Nav (magenta...magenta), anti-CASPR (green), and anti-Kv2.1 (cyan). (B) Cerebellum labeled with anti-GABA-AR β1 (white...white) and anti-GABA-AR β3 (green). (C) Hippocampus labeled with anti-Kv2.1 (magenta), anti-AnkyrinG (green... secondary antibodies will bind to each primary antibody.      Look for primary antibodies that have ... combination of antibodies:   Target (Rat) Primary antibody Secondary antibody SCN1A sodium...channels anti-pan-Nav R-mAb K58/35R (Mouse IgG2a) Goat anti-mouse IgG2a CASPR anti-CASPR mAb ... Goat anti-mouse IgG1 Kv2.1 channel anti-Kv2.1 KC (Rabbit IgG polyclonal) Goat anti-Rabbit IgG...

...Addgene blog Antibodies 101: Introduction to AntibodiesAntibodies 101: IsotypesAntibodies 101: Chimeric...secondary antibodies to bind. We also know that changing the Fc domain can impact an antibody's binding ... affect antibody applications like ELISAs or Western blots, they are key in helping antibodies drive an...responses? We got you! You can also review antibodies in our Antibodies 101 animation.)  Many types of immune...receptors that bind to the Fc domain of an antigen-bound antibody and induce (or, in the case of the Fc receptors...a viral infection a) antibody-dependent complement deposition (ADCD) b) antibody-dependent cellular cytotoxicity... et al., 2019.    Antibody-dependent complement deposition (ADCD) Antibodies can bind to a pathogen...

...Addgene.org Addgene’s General Use Antibodies Antibody guide Antibody protocols Additional resources ...of finding a ‘good’ antibody is something we’ve all experienced. Finding an antibody that works for your... common tags, the antibodies used to detect them, and how they can fix your antibody woes.   What are...without the need for an antibody. Tags can enable the separation of detection of a mutant protein vs. endogenous...monoclonal antibodies. Below we review the most commonly available tags and their antibodies along with...Addgene blog Antibodies 101: Epitope availability Plasmids 101: Protein Tags Antibodies: Of Myc and Men...Epitope tags eliminate the need for target-specific antibodies and have been widely used across species and ...

...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...

...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 ...

...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...

... validated antibody, we recommend using antibody search engines and antibody data repositories to find... that can help: Antibody data repositories, which share validation data, and antibody search engines, ...many different antibody catalogs at the same time.  If you're looking for an antibody, especially a validated...improving antibody quality throughout research, consider checking out the Only Good Antibodies community...If you’re looking for an antibody that works for your experiment, you may be overwhelmed by options —...find an antibody for your experiment. Search engines let you easily find and compare available antibodies...and experimental data to help you decide if the antibody is a good fit for your experiment. They come in...

...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...

...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...  Once you get your exciting new antibody, the second thing you’ll want to do is store it away safely. ... check it whenever you use the antibody. This information is important, and you’ll need to track it throughout...accidentally leave your antibody out on the bench overnight. Do not panic! Typically, antibodies are okay if they...light-sensitive conjugate on the antibody left exposed to light?  Has this antibody been left out, gone through...are planning to conjugate your antibody in the lab, you’ll therefore want to research your conjugation ...

... IPI anti-integrin toolset IPI’s anti-integrin toolset comprises 26 recombinant antibodies (rAbs) either...Addgene's Antibody Guide Antibodies 101: Flow Cytometry Antibodies 101: Introduction to Antibodies...Table 1: IPI antibodies  Antibody Target Proteins Reactivity Isotype Source Species Anti-Integrin...unique antibodies that bind discrete subunits or particular subunit pairs. Some of the antibodies in the...property in an antibody is extremely rare. While many function-blocking integrin antibodies exist, almost...are chimeric antibodies composed of rat or mouse variable regions fused with rabbit constant domains. All...monoclonals, we anticipate that our chemically identical hybridoma-derived antibodies will also work in...

...license.    In the third plot, anti-CD14 and anti-HLA-DR antibodies conjugated to fluorescent markers...Cytometry Plasmid-based Recombinant Monoclonal Antibodies Introduction to Antibodies  ... is a cell, but it could be debris or unbound antibodies.) Each event read by the machine is plotted as...conceptualize where the populations are, try drawing a quadrant over the plot that divides it into four, using...different populations by imagining (or drawing) a quadrant over the dot plot. Created with biorender.com....Biorender! While a helpful visualization tool, a quadrant will often be too clean for real-world data. Most...using a panel of markers, which is comprised of antibodies conjugated to fluorescent markers and/or cellular...

...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...

...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...

...and detection with antisera: a method for studying antibody specificity and antigen structure. Proceedings... repeatedly stripped of an antibody and reprobed with different antibodies allowing multiple targets to...binding to the antibodies during the stain. The ideal blocking buffer depends on the antibodies and labels...your primary antibody Once the membrane has been blocked, it is stained with a primary antibody against your...interest. But which antibody should you use? You will likely find several different antibody options targeting... their antibody binds to protein of the correct size. Many vendors will show that the antibody binds to...that the antibody efficiently binds to endogenously expressed protein at physiologically relevant levels...

...should first note that quantitative PCR (qPCR) has been a powerful tool for quantifying AAV for some time ...producing AAV vector preparations, we have used quantitative PCR to evaluate the physical titer of our preps...digital PCR (ddPCR) comes in. AAV titering using quantitative PCR Before we dive into the details of ddPCR...of a standard - a set of reactions with known quantities of DNA. Additionally, an AAV reference, a virus...by ddPCR begins with diluting the virus. It is important to note that the dynamic range of the ddPCR is...PCR plate. Maintaining a high droplet count is important for calculating the concentration of your sample...separation (Witte et al., 2016). This is particularly important if your amplicon is longer than 150 base pairs...

...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:...

...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...