...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...

...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 ...

...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 ...

...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-...

...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...

...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 ...

...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:...