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  1. Synthetic Biology - Overview

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    Collection
    ... Standard European Vector Architecture Open Plant - Plant SynBio resource EraSynBio - The EU/USA SynBio...Mukhopadhyay Richard Murray Erin O'Shea Martin Parniske Antonio Richart Herbert Sauro Claudia Schmidt-Dannert Pamela...pairs. Browse Plasmids by Model Organism Bacterial Plant Yeast Mammalian Worm Fungal Algal Featured SynBio...FREQ-Seq Kit FX Cloning Kit GreenGate Cloning System (Plant) MoClo Modular Cloning Modular PUF Assembly Kit ...Kit Optogenetics Pertz Lab cpFRET Kit Rinehart Recombinant Phosphoprotein Kit S. cerevisiae Advanced Gateway...
  2. AAV for Neuronal Tracing

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    Collection
    ...pAAV-synP-FLEX-splitTVA-EGFP-B19G Can be used to complement deletion-mutant rabies virus AAV1 Wickersham 100798 pAAV-syn-FLEX-splitTVA-EGFP-tTA...AAV together can be used to complement deletion-mutant rabies virus AAV1 Wickersham 100799 pAAV-TREtight-mTagBFP2...the cell population of interest with a deletion-mutant tracing virus that lacks the ability to infect ... To enable monosynaptic spread of this deletion-mutant rabies, the essential G protein is provided to ...2007a. Retrograde neuronal tracing with a deletion-mutant rabies virus. Nat Methods. 4(1):47-9. PMID: 17179932... viral prep format. Please note this does not guarantee viral vector packaging service, but lets us know...
  3. 6 Tips for Grant Writing

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    Blog Post
    ...improve your grant writing and hopefully increase your success rate. 1. Submit your grant early! First... 2. Originality is essential for grant success The two most important things reviewers will be looking...in writing small travel grants to attend conferences and a seed funding grant for a proof-of-principal...ones. With grant applications there’s going to be an element of luck involved so it’s important not to throw...www.assaygenie.com. No matter what facet of academia you are in, grant writing can be the lynch pin that results in your...First off, and perhaps the most important bit of advice I can suggest, is to give yourself plenty of time..., give it back to you, and for you to make the relevant alterations before submitting - handing them your...
  4. The Twelve Days of Antibodies

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    Blog Post
    ...online Antibody Guide!    🎶On the fourth day of Antibody, Addgene gave to me…  A recombinant Anti-Arl13b...me… A recombinant Anti-PSD-95!    🎶On the seventh day of Antibody, Addgene gave to me… Antibodies developed...day of Antibody, Addgene gave to me…  A recombinant Anti-DYKDDDDK!   🎶On the ninth day of Antibody, Addgene...neuroscience antibody collection!   🎶On the tenth day of Antibody, Addgene gave to me…  A recombinant Anti-APP...day of Antibody, Addgene gave to me…  An Antibodies 101 eBook!  🎶On the second day of Antibody, Addgene...Addgene gave to me…  A recombinant anti-GFP!    🎶On the third day of Antibody, Addgene gave to me…  An online... of Antibody, Addgene gave to me… The Addgene Antibody Data Hub!    🎶On the sixth day of Antibody, Addgene...
  5. Antibodies 101: Conjugation

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    Blog Post
    ...commercially conjugated antibodies available, that never guarantees the antibody you need will be conjugated... the methods for antibody conjugation can be quite variable from antibody to antibody or batch to batch...research antibodies. Lysine residues are found throughout antibody sequences, up to 80 per antibody (Mueller...abundance targets with significant multiplexing potential. You will find antibody-oligonucleotide conjugates... conjugate antibodies directly to beads for purification purposes.  Coupling up - antibody conjugation...mechanisms. Most take advantage of chemical groups intrinsically present on antibodies, such as the primary...disulfide bonds between antibody chains. 3) Endogenous carbohydrates added to antibodies post-translationally...
  6. New Antibody Animation!

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    Blog Post
    ...about antibodies? Our latest animation is here to help! Join Abi as they explain what antibodies are, ...role in the immune response, and the different antibody isotypes found in mammals.      Subscribe to...
  7. Antibodies 101: Validation

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    Blog Post
    ...does not guarantee recognition of the target antigen. You want to make sure that your antibody binds to... a great antibody signal in the non-mutant line but no signal in the mutant line! Recombinant Strategies... an antibodyAntibodies and Reproducibility First, let’s review why validation is so important. Antibodies...expression of antibody-independent approaches to your antibody-dependent approach. Antibody-independent ...by your antibody. You perform immunocytochemistry on these mutant cells as well as a non-mutant line and...means, such as through antibody databases like Antibodypedia or through your antibody provider(s) if they...choose an antibody? Check out our Selecting an Antibody blog post! Learn more about Antibodies and their...
  8. Antibodies 101: ChIP

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    Blog Post
    ...s blog Antibodies 101: Immunoprecipitation  Antibodies 101: Polyclonal Antibodies Antibodies 101: Introduction...DNA Incubate your antibody, or antibodies, with your samples, and then use an antibody capture method such...Introduction to Antibodies Resources on Addgene.org Find plasmids encoding recombinant antibodies at Addgene... ChIP works by using antibodies to capture protein:DNA complexes with antibodies specific to your protein...mask the epitopes antibodies recognize. It is therefore best to either select antibodies validated for ChIP...incubate the sample with your antibody.  Both polyclonal and monoclonal antibodies can work well in ChIP. If...your antibody:protein:DNA complexes and finally (5) isolated the DNA from your antibody:protein:DNA complexes...
  9. Antibodies 101: Immunoprecipitation

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    Blog Post
    ...primary antibody interfering with downstream assays. This is particularly advantageous if the antibody used...specificity of anti-epitope antibodies make this approach appealing, but the disadvantages must be carefully...Addgene's antibody posts  Resources on Addgene.org: Find plasmids encoding recombinant antibodies at ... the same basic strategy: immobilize the antibody or antibody-target protein immune complexes, wash away... this method, the free antibody is incubated with the protein mix and antibody-target protein immune complexes... the process of anchoring an antibody, often called the capture antibody, to agarose or magnetic beads...and isotype of their primary antibody. To circumvent this, a recombinant protein, Protein A/G, was developed...
  10. Antibodies 101: Isotypes

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    Blog Post
    ...Why are antibody isotypes important? To your immune system, an antibody’s isotype is important because...by its heavy-chain constant region (see Antibodies 101: Introduction to Antibodies for a refresher). There...Transportation of antibodies to places they can’t reach on their own. The antibody’s heavy-chain constant domains... the same constant heavy chain domain as your primary antibody but doesn’t bind your antigen of interest...valency of 10 antigens. An extra constant domain replaces the hinge region in IgM antibodies. IgM has no...blog Learn about the advantages and disadvantages of using monoclonal antibodies Read about other techniques...that you can use antibodies for in our Intro to Antibodies post Check out our Antibodies 101: What is an...
  11. SARS-CoV-2 Pseudotyped Virus

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    Collection
    ...information. Plasmids Expressing Spike and Spike Variants Full-length wildtype SARS-CoV-2 spike protein ...D614G change which is present in many circulating variants. The table below lists expression plasmids that...protein (MERS, CoV-1, CoV-2), modification, or variant. pBOB-CAG-SARS-CoV2-Spike-HA (Addgene plasmid #141347...Addgene Blog Posts Viral Vectors 101: Pseudotyping Antibody Neutralization Response Against Pseudoviruses ...Pseudoviruses Expressing SARS-CoV-2 Spike Protein Variants Addgene COVID-19 Resources and Collections COVID-19 and...
  12. EXtracellular Plasmid RESource (EXPRESs) Consortium

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    Collection
    ...surface receptor and secreted proteins as soluble recombinant molecules....surface receptor and secreted proteins as soluble recombinant molecules. The proteins are typically used in...identify novel interactions or pathways that are important in cellular recognition processes. Laboratories...negative controls Zebrafish receptors Zebrafish antibody plasmids P. falciparum merozoite surface protein...invasion by Plasmodium falciparum. Crosnier C, Bustamante LY, Bartholdson SJ, Bei AK, Theron M, Uchikawa...
  13. Adeno-associated virus (AAV) Plasmids

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    Collection
    ...originally discovered as contaminants of adenovirus stocks. One major advantage of using AAV for research...low immune response and low toxicity. Although recombinant AAV does not integrate into the host genome, ...AAV Viral integration AAV serotypes Pseudotyping Want to save time? Check out our Viral Vector Packaging...129024 pProvector_AAV2_586_PLGLAR AAV2_586_PLGLAR Recombinant AAV2 rep/cap plasmid modified to include an 18...Gradinaru 212708 pAAV SCH9 SCH9 A SCHEMA evolved AAV variant effective at targeting subventricular NSCs Schaffer...
  14. Retrovirus Plasmids

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    Collection
    ...Large T antigen for creation of immortalized cells (see Weinberg Lab page for resistance variants) Weinberg...results in long-term transgene expression. Some disadvantages of γ-retroviruses are that they can only transduce... Plasmids Common γ-retroviral transfer plasmids. Want a retroviral plasmid with your gene of interest?...expression; also expresses GFP. Also see pMIG-w , a variant that has WRE, a post-transcriptional regulatory...
  15. CRISPR Plasmids - Drosophila

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    Collection
    ...Double nicking strategies reduce unwanted off-target effects. Nickase mutants can also be used with a repair... gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish Xenopus CRISPR Resources...Promoter PI Publication Nick CRISPR/Cas nickase mutants introduce gRNA-targeted single-strand breaks in...created by wild type Cas enzymes. To use a nickase mutant, you will need two gRNAs that target opposite strands...
  16. CRISPR Pooled gRNA Libraries

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    Collection
    ...collection differ based on some important criteria, and it is important to know which library best... gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish Xenopus CRISPR Resources...with the pooled library to create a population of mutant cells that are then screened for a phenotype of...representation. CRISPR screening experiments require large quantities of cells and therefore, may not be suitable for...best suits your particular experiment. Some important variables include: Type of genetic modification - ...
  17. Luminescent Imaging with Nano-lanterns

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    Blog Post
    ... 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...
  18. Antibodies 101: Affinity Tags

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    Blog Post
    ...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 ...
  19. Antibodies 101: Multiplex Immunofluorescence

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    Blog Post
    ...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...
  20. Antibodies 101: Fc Effector Functions

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    Blog Post
    ...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...
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