... create a fusion with a reporter protein such as GFP? All of these outcomes can be most effectively achieved...conjunction with activator or repressor domains to control gene expression. Base editors can help you make...add a tag, you will also need to deliver a donor plasmid or single-stranded DNA oligonucleotide, so you ...typically used for gRNA. May contain reporter gene (e.g. GFP) to identify and enrich positive cells or a selection...transfer vectors. May contain reporter gene (e.g. GFP) to identify and enrich positive cells. Likely contains...generate stable cell lines. Packaging and Envelope plasmids provide the necessary components to make lentiviral...for in vivo viral delivery. Cas9 mRNA and gRNA Plasmids containing gRNA and Cas9 are used in in vitro ...

...transformation Read our Plasmids 101 blog posts Resources on Addgene.org Browse our Plant Plasmids and Resources...page Find synthetic biology plant plasmids Find CRISPR plant plasmids ...its function. In nature Agrobacterium contains a plasmid with a T-DNA (transfer-DNA) region. During infection...check for GUS-like activity of your tissue. Run a control with Agrobacterium lacking the GUS gene since plants...promoter activity, for example. Useful positive controls could include: pSB96 - pL2_pSB90_2x35S::fLUC-I...tMAS::rLUC-I::pMAS. Fluorescent proteins (FP, e.g. GFP) can be used in many ways. Often a FP is fused to...localization of your protein of interest. Find all plasmids from the Lee-Parsons lab. Test different leaf...

... is now complete! pAAV-CAG-GFP (plasmid 37825) and pAAV-hSyn-EGFP (plasmid 50465) are now available as...continue to add more controls, and have recently released these controls: pAAV-hSyn-DIO-EGFP (50457-AAV5): ...sensors Optogenetics Recombinases And more! Control AAV For many AAV experiments, we recommend validating...entire collection! We are expanding our entire control AAV inventory to give scientists access to a wide... Email us or check our website to access them! Controls pAAV-hSyn-hChR2(H134R)-EYFP (26973-AAVrg) pAAV-hSyn-hChR2...our entire AAV inventory. Our new AAVs include: EGFP-expressing AAV for serotype testing Calcium sensors... testing AAV, which are small (20 ul) samples of EGFP-expressing AAV packaged in various serotypes. This...

...conjugative mobilization of broad host-range IncQ plasmids." Plasmid 62.2 (2009): 57-70. PubMed PMID: 19465049...page Deposit microbiology plasmids with Addgene Find synthetic biology plasmids ...family-RP4 conjugation system. This system can deliver plasmids to both Gram-positive and Gram-negative cells.... Wang lab further developed a modular suite of plasmids (pGT) with various replicative origins that can...selectable marker and a desired genetic payload (ex: GFP). Thus, cells can be both sorted using FACS and isolated...genera across four phyla that have taken up the plasmid through conjugation in vivo studies. However, they... tools for microbiome studies.  Find the MAGIC plasmids here Paper-based synthetic biology approach to...

...protein control. We also added the toxin-antitoxin stability cassette Axe-Txe so that the plasmid could ...accessory genes gvpNJKFGWV marked with Emerald GFP. These plasmids can be used directly in transient transfection... 1248 ng of CMV-gvpA-IRES-BFP plasmid and 954 ng CMV-gvpNJKFGWV plasmid into 70%-confluent well of HEK...in the absence of antibiotics, resulting in the plasmid pBAD-bARGSer-AxeTxe (Fig. 1a).  With pBAD-bARGSer-AxeTxe... Fig. 1: Expressing bARGSer in E. coli. (a) Plasmid diagram of pBAD-bARGSer-AxeTxe. (b) Methods to ...overexpressed compared to accessory proteins. Excess gvpA plasmid added into the transfection mixture in HEK293T ...mammalian species to produce gas vesicles. New mARG plasmids Our new mARGs come as a set of two PiggyBac integratable...

...Addgene.org Find plasmids from the Bethune Lab Find Plasmids from the Roux Lab Find Plasmids for FRET ... No spatiotemporal control. Many PPI detection methods lack spatiotemporal control or an “on/off switch...partners if they are enriched relative to controls using Split-BioID-GFP and BioID run on six unrelated proteins... by creating Split-BioID, a spatiotemporally controllable version of the proximity-dependent biotinylation...method Enter Split-BioID, a spatiotemporally controllable version of the BioID protocol. The critical ...BirA* fusion proteins are expressed in cells via plasmids or by the creation of stable cell lines and these...methods, including BioID, is its spatiotemporal controlled labeling of interacting proteins; nearby proteins...

...pAAV-CAG-GFP CAG GFP Control Boyden 50465 pAAV-hSyn-EGFP hSyn EGFP Control Roth 50469 pAAV-CaMKIIa-EGFP CamKIIa... 135631 pAAV-S5E2-GFP-fGFP E2 regulatory element GFP, membrane-targeted GFP Control Dimidschstein 192552...pAAV-CAG-eYFP CAG EYFP Control Gradinaru 104061 CAG-NLS-GFP CAG NLS-GFP Control Gradinaru 105530 pAAV.CMV.PI....CamKIIa EGFP Control Roth 59462 pAAV-CAG-tdTomato CAG tdTomato Control Boyden 83900 pAAV-mDlx-GFP-Fishell...pAAV-mDlx-GFP-Fishell-1 Dlx GFP Control Fishell 99130 pAAV-mDlx-NLS-mRuby2 Dlx mRuby2 Control Gradinaru 104055 pAAV-CAG-...Promoter Description Category PI 37825 pAAV-CAG-GFP CAG GFP Control Boyden MaCPNS2 These viral vector preparations...Promoter Description Category PI 37825 pAAV-CAG-GFP CAG GFP Control Boyden CAP-B10 These viral vector preparations...

...In a previous post from our Plasmids 101 series, we learned how the Cre-loxP recombination system can... even in the presence of Cre recombinase. This plasmid now specifically drives expression of mCherry instead... should contain a reporter DNA sequence (such as GFP) in the sense orientation, followed by coding sequence...transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic reporter would ensure...et al. Nat Biotech 2003): In the presence of Cre, GFP expression is lost, allowing for rescued expression...2286602. PubMed Central PMCID: PMC3406316. Find FLEx plasmids from these Addgene depositors: Ed Boyden Bernardo... plasmids...

...under the control of the mDlx enhancer element Gordon Fishell 83900-AAV1 pAAV-mDlx-GFP-Fishell-1 GFP expression...under the control of the mDlx enhancer element Gordon Fishell 83900-AAV2 pAAV-mDlx-GFP-Fishell-1 GFP expression...under the control of the mDlx enhancer element Gordon Fishell 83900-AAV5 pAAV-mDlx-GFP-Fishell-1 GFP expression...under the control of the mDlx enhancer element Gordon Fishell 83900-AAV8 pAAV-mDlx-GFP-Fishell-1 GFP expression...under the control of the mDlx enhancer element Gordon Fishell 83900-AAV9 pAAV-mDlx-GFP-Fishell-1 GFP expression... the control of the mDlx enhancer element Gordon Fishell 83900-AAVrg pAAV-mDlx-GFP-Fishell-1 GFP expression... the control of the mDlx enhancer element Gordon Fishell 83900-PHPeB pAAV-mDlx-GFP-Fishell-1 GFP expression...

...you find what you need.   More Plasmid Resources from Addgene: Plasmids 101 blog series Addgene's Molecular...function of Your Gene Of Interest (YGOI) in a controlled environment under various conditions. The first...need. You will also have to make sure that your plasmid has been incorporated into the host organism, usually...great for overexpressing YGOI in an isolated, controlled environment. In most cases bacterial expression...open reading frame (ORF) of interest –  try pRS plasmid shuttle vectors. You may prefer the more traditional...using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. pPD80_08) and following the expression pattern...details and guidance for choosing your perfect plasmid backbone. Or just ask –  leave a message in the...

...Fluorescent Proteins Control Protein Expression with Cre-lox Control Protein Activity with GFP Resources at ... the plasmids from Zhou et al here. In addition, it could be possible to use pdDronpa to control other...transmitted by intermediate messengers. Optical control of kinase activity can provide greater spatiotemporal...There is no common method for creating optically controlled kinases. The Solution: In Zhou et al, the Lin... the delayed effects because you can precisely control the timing of kinase activity. 2. Studying negative...photoswitchable kinases in vivo not only allows for precise control of a signaling pathway in the context of a whole...Pengpeng Li, Kang Shen, and Michael Z. Lin. "Optical control of cell signaling by single-chain photoswitchable...

...without mCherry and GFP fluorescent controls. Alternatively, you can order individual plasmids from the library...Addgene.org Plasmid Pooled Libraries Pooled library amplification Zhang Lab CRISPR Plasmids The Human TFome...streamline identification. While the Zhang lab controlled their multiplicity of infection so that a single...disease depends on both genetic and expression control systems to mimic illness on a molecular level. ... of the collection, we’re releasing individual plasmids in batches. If there’s something you’d like that...

...antibodies against popular fluorescent proteins (GFP and YFP) and protein tags (c-MYC and HA). We have...materials, repositories provide standardized quality-control and technical support to the community. Thanks ...about Scientific Sharing: The 10 Most Distributed Plasmid Technologies in Addgene's First 10 Years Data...

...curated collections of plasmids as kits with greatly reduced costs per plasmid. We generally create kits...just the tool you need. The plasmids in this portion of the BIOFAB Plasmid Set contain terminators of ...populations. This kit consists of a 48 plasmid adapter library, with each plasmid carrying a unique barcoded Illumina-M13F...researcher at a nonprofit institution, you can order plasmids covering a wide range of fields from Addgene. ...kits to allow you to easily use a set of plasmids together on a tight budget. To help popularize our ...order from the repository: FX Cloning Kit BIOFAB Plasmid Set pCri System Cell Free Protein Expression Kit... production (a variety of pExpression vectors) plasmids. The type IIS restriction sites used in these ...

...pAAV-mDlx-GFP-Fishell-1 Dlx GFP Control Fishell 83895 pAAV-hDlx-Flex-GFP-Fishell_6 Dlx GFP Control Fishell...AAV-CAG-GFP CAG GFP Control Boyden 51502 AAV pCAG-FLEX-EGFP-WPRE CAG EGFP, Cre-dependent Control Zeng 50465...50465 pAAV-hSyn-EGFP Syn EGFP Control Roth 50469 pAAV-CaMKIIa-EGFP CamKII EGFP Control Roth 114472 pAAV-hSyn-mCherry...Cre-dependent Control Roth 59462 pAAV-CAG-tdTomato CAG tdTomato Control Boyden 50457 pAAV-hSyn-DIO-EGFP Syn EGFP...quality control, including AAV titering by SYBR green qPCR , confirmation of the transfer plasmid with PCR...pAAV-hSyn-mCherry Syn mCherry Control Deisseroth 114469 pAAV-CaMKIIa-mCherry CamKII mCherry Control Deisseroth 50459...Cre-dependent Control Roth 55650 pAAV-hSyn Con/Fon EYFP Syn EYFP, Cre and Flp-dependent Control Deisseroth...

...right negative control is critical to the success of BiFC experiments. The ideal control is a mutated variant...-to-transfect cell lines where plasmid dosage can be tightly controlled. This is also theoretically possible...because it relies on a single BiFC pair. Find BiFC plasmids for your experiment at Addgene! How to Design ... work. You will also need to generate negative control constructs — more on this later. For more on constructing...check out our article: Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection. Cellular... measure the fluorescence of a single FP. Your control construct may generate a modest level of fluorescence...intensity of your BiFC constructs to your negative control to determine whether your PIPs actually do interact...

...blot showing GFP expression in cells that have been transfected with a plasmid encoding GFP. Actin, a common... be used for. Addgene’s Antibody Plasmid Collection includes plasmids that express antibodies, nanobodies...immunoglobulin genes, recombinant antibodies are plasmid-based. Their sequences can be optimized for specificity...protein (Wang et al., 2013). These proteins are also plasmid-derived. However, a disadvantage to ScFv is their...nanobodies, or synbodies, can be produced from plasmids, saving time and money. Nanobodies and synbodies...detection. You’ll also want to use the appropriate controls for your experiment to account for potential non-specific...quantitatively if you also load the gel with appropriate controls for normalizing the protein concentration. To ...

... from the default GFP (green) to RFP, CFP, or YFP fluorescence. In Brainbow 2.1, GFP is tagged with a ...and activate fluorescence of the downstream OFP, GFP, or RFP. Cells that express the construct but do ...use in Drosophila melanogaster and is under the control of Flp recombinase. The XFPs in Flybow are membrane-tethered...a specific tissue. XFPs are introduced using a plasmid construct called Zebrabow-GateDest containing an...generates cells that randomly express either nuclear GFP, membrane CFP, cytoplasmic YFP, or cytoplasmic RFP...introduced in vivo either by electroporation of plasmids, or using AAV vectors. The Imai lab also developed...incorporates the highly photostable fluorophores mOrange2, EGFP, and mKate2 (red) to solve the problematically low...

...Using an epifluorescence microscope and GFP fusion proteins controlled by promoters specific to the immune... to track where GFP labelled B cells were in mounted slices of these organs. These GFP labelled cells ...enabled by fluorescent proteins The optimization of GFP and its derivatives by the late Roger Tsien changed...immune lineage, scientists could easily track GFP expressing cells. For example, by harvesting lymph nodes... the infiltration of neutrophils expressing LysM-GFP (green) into a draining lymph node after infection...researcher can flank fluorescent proteins in a plasmid with loxP sites such that their expression is turned...Check out Our Fluorescent Protein Guide Pages Find Plasmids for Subcellular Localization Find Fluorescent ...

...hiPSCs  EMX1  67% Mouse BM cells from GFP transgenic mouse  GFP  ~60-65% Human hepatocytes  Myd88  ~...infiltrate cells. Options for CRISPR delivery include plasmid transfection, RNP electroporation, and viral transduction...types they tested. Interested in using Nanoblades? Plasmids are available from Addgene. Genome editing primary...to DNA transfection of gRNA and Cas9 expression plasmids. These results are in line with previous research...editing in their livers, while non-injected and control gRNA injected mice had no detectable editing. Additionally...out our comprehensive CRISPR guide Find CRISPR plasmids and resources Learn about viral production ...

...pAAV.GfaABC1D.PI.Lck-GFP.SV40 Control Baljit Khakh AV-5-PV2380 100896-AAV5 pAAV.GFA104.PI.eGFP.WPRE.bGH Control Philip...pCAG-FLEX-EGFP-WPRE Control Hongkui Zeng AV-9-PV0101 105530-AAV9 pAAV.CMV.PI.EGFP.WPRE.bGH Control James ...pAAV-Ef1a-DIO EYFP Control Karl Deisseroth AV-1-ALL854 51502-AAV1 AAV pCAG-FLEX-EGFP-WPRE Control Hongkui Zeng....SV40 Control Loren Looger AV-1-PV3661 98927-AAV1 pENN.AAV.CAG.Flex.GFPsm_myc.WPRE.SV40 Control Loren ...pAAV-Ef1a-DIO EYFP Control Karl Deisseroth AV-2-ALL854 51502-AAV2 AAV pCAG-FLEX-EGFP-WPRE Control Hongkui Zeng...pAAV.CMV.LacZ.bGH Control James M. Wilson AV-5-PV1917 105541-AAV5 pENN.AAV.CamKII0.4.eGFP.WPRE.rBG Control James ...pENN.AAV.CMVs.TurboRFP.WPRE.RBG Control James M. Wilson AV-5-PV2213 105547-AAV5 pENN.AAV.EF1a.eGFP.WPRE.rBG Control James M. ...

...fluorescent protein reporter under the control of promoters that control cell cycle gene expression.  Additional...-up approach will allow me to create a fully controllable and reproducible set up,” Bernach explains. ...containing unstable fluorescent proteins under the control of promoters responsive to water or nutrient levels...fluorescent protein topics page Get the basics on GFP Find microbiology blog posts Resources on Addgene.org...Addgene.org Find fluorescent protein biosensors Browse Plasmids from the Tsien lab Browse the Michael Davidson...variant library is monitored by fusing the variants to eGFP followed by an internal ribosomal entry site and...a variant has wild-type levels of abundance, the eGFP will be stable and fluoresce. But if the variant...

...from the Church lab uses this system to control expression of GFP.)  To turn this inducible system into ...Promoters control the binding of RNA polymerase and transcription factors. Since the promoter region ...promoters can be regulated via positive or negative control. Positive Repressible: An activator protein is ...system, Gossen and Bujard created the tetracycline-controlled transactivator (tTA). They fused TetR with the...Tet-Off system.  For more information on Addgene Tet plasmids, see our Tetracycline resource page. Cumate Switch...interest to be expressed. These permit exquisite control of gene expression and tracing of gene expression...that sometimes it is necessary or desirable to control or look at multiple genes at the same time.  The...

...stability of modified Ti plasmids during plant infection, as well as plasmid yield during preparation ...GreenGate Plasmids Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More Plasmids 101: Origin...Agrobacterium carries a “tumor-inducing” or Ti plasmid, which allows it to transfer genetic material into...in two species: Agrobacterium and E. coli. Like plasmids used in other systems, binary vectors are cloned...It’s a lot of information to fit into a single plasmid, so most first-generation binary vectors are quite...Arabidopsis, and corn. For both β-glucuronidase (GUS) and GFP reporters, as well as multiple transient and transgenic.... The kanamycin antibiotic resistance gene is controlled by a weaker constitutive Nos promoter, and it...

...Reporter Gene like Luciferase and GFP Resources on Addgene.org Find Plasmids for Your Research     ...Once you've gotten to know what a plasmid is in general, it's time to take a look at some of its parts...origin of replication are necessary to replicate plasmids and make sure the cells maintains it. Here, we'll...the promoter, which is necessary for getting the plasmid express our gene of interest. This element is responsible...target genes. The sequence of the promoter region controls the binding of the RNA polymerase and transcription.... This means that the promoter present in your plasmid backbone must to be compatible with the type of...transcript, you will also need to make sure your plasmid has a promoter suited to working in your host organism...

...Packaging Service AAV Controls Viral Vector Packaging Service: Control AAV Browse our control AAV that express...CAG-NLS-GFP CAG NLS-GFP Constitutive PHPeB Gradinaru 105530 pAAV.CMV.PI.EGFP.WPRE.bGH CMV EGFP Constitutive...quality AAV preps from select plasmids in the repository. Browse our control AAV collection, including fluorescent...Fluorophore/Tag Activity Serotype PI 37825 AAV-CAG-GFP CAG GFP Constitutive 1, 2, 5, 8, 9, rg*, PHPeB, CAP-B10...*, PHP.eB, PHP.S Boyden 83900 pAAV-mDlx-GFP-Fishell-1 Dlx GFP Constitutive 1, 2, 5, 8, 9, rg*, PHP.eB ...dependent PHP.V1 Gradinaru 83895 pAAV-hDlx-Flex-GFP-Fishell_6 Dlx GFP Cre dependent 1, 2, 8, 9, rg* Fishell 83894...Browse all Control AAV *AAVrg = retrograde serotype, produced with the AAV retro helper plasmid from the...

...virus (MoMLV) and consisted of two plasmids: the packaging plasmid containing all the structural genes...crossover event between the vector plasmid and the packaging plasmid caused the Psi sequence to be placed...expression of the transgene   Transfer plasmids The transfer plasmid is the vector you clone your gene of...usually 293T cells, with your transfer plasmid and your packaging plasmids. For this, you can use the transfection...containing the gene of interest (pTet-IRES-EGFP, pPRIME-Tet-GFP-FF3) and one with either tTA or rtTA (FUW-M2rtTA...techniques are still used and efficiently transfer plasmid DNA efficiently into dividing immortalized cells... (Gag, Pol, Tat, Rev and Env) and the transfer plasmid encoding the insert of interest as well as sequences...

...cargo in one plasmid (the transfer plasmid), the envelope (Env) gene on a separate plasmid (envelope plasmid... One such control is dividing up the essential components of a virus into different plasmids. For a retroviral...Gag structural protein on yet a third plasmid (packaging plasmid). Viral production then proceeds by transfection...virus and its infectivity in check. Engineering controls, safety systems that are built into how something... transfection of all three plasmids into a producer cell line, such as HEK-293T, production of viral particles... and Pol genes are not encoded in the transfer plasmid, the component that physically integrates into ... retroviral packaging components onto distinct plasmids for safety.    Viral cargo-specific safety...

...of the new plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. ... blot for the myc-tagged protein GFP-smFP-myc expressed from Plasmid 98926. See the Applications section...Think we missed something hot? You can pitch a Hot Plasmids section (antibodies and viral preps welcome!) ... you can also find several other constructs in plasmid form. Fig. 1: Summary of the approach and...JEDI-2p viral preps here (and here!)Find JEDI-2p plasmids here!  Liu, Zhuohe et al. Sustained deep-tissue...represent a long-sought tool for researchers to control K+ gradients and can be used as a potent optogenetic... improve the performance of HcKCRs. Find HcKCR plasmids here!   Govorunova EG, et al. Kalium channelrhodopsins...

...Browse Tetracycline Plasmids Tet-regulated Lentiviral RNAi Vectors Curated Plasmid Collections Background...interest tTA expression plasmid Components for a tetracycline on system are: Plasmid containing a tetracycline-dependent...generation lentiviral plasmid for inducible expression of shRNA; neomycin selection; plasmid 21915 has puromycin...option rtTA On Kowarz 16623 pBI-GFP Expression of your gene of interest & GFP from a bidirectional tet-responsive...and an inducible promoter at its 3' end, which controls GFP expression tTA Off Verma 26803 pEnt L1L3 EF1a-tTA...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...pTet-IRES-EGFP Lentiviral plasmid for inducible expression of transgene of interest and EGFP None Either...

...spatiotemporal control over labeling methods as compared to transgenic animals (17).  These viral plasmids are available...mouse brains expressing green fluorescent protein (GFP) (6).   The Dodt and Frank Bradke laboratories later...subpopulation of neurons are brightly labeled with GFP (e.g. Thy1-YFP-H mice).  However, we recently developed...University of Miami Miller School of Medicine. Find plasmids from the Lee Lab here. Pantelis Tsoulfas is an...University of Miami Miller School of Medicine. Find plasmids from the Tsoulfas lab here.     References: H... nerve afferents labelled with a AAV8 expressing eGFP under the human ubiquitin C promoter. Anatomical...

...their Cas9 and gRNA backbone plasmids with Addgene. Additional control plasmids for the TKO library will ...Neurosci 2016. PubMed PMID: 26950006 Plasmid ID Plasmid Name Plasmid Type 74309 pcDNA3.0-TRPV4-p2A-ferritin-p2A-mCherry...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These brief articles...features and applications of a partiular plasmid or set of plasmids. We hope that these articles make it ... you to find and use the plasmids you need. You can find all the hot plasmids from 2016 below. With over...over 45,000 plasmids, we can't write posts for every great plasmid that comes into the repository, but...write about your plasmids in a future blog post. No time to read? Listen to our hot plasmids segment on the...

...the transfer plasmid, a plasmid encoding Rep and serotype-specific Cap genes, and a plasmid encoding adenoviral...Production Titer Quality Control Resources Lentivirus Production Titer Quality Control Resources Overview of... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...subunit (RPP30) as a control for normalization purposes are being used. Quality Control Mycoplasma The 293T... coli . In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits. Resources...Addgene's viral production methods and quality control for lentivirus and AAV. Viral...virus preps are titered and subjected to quality control by Addgene before being distributed to customers...

...Purpose PI 17446 pLenti CMV GFP Hygro (656-4) GFP Hygromycin 3rd gen lentiviral eGFP expression vector, CMV...Pooled Barcoding Libraries Control Addgene's lentiviruses are made from plasmids that are currently in our... list of gRNA plasmids targeting specific genes. For more information on CRISPR plasmids and function,... 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A GFP Zhang 61425 ...library expresses EGFP for easy visualization via direct fluorescence. Morris Control Lentivirus Browse...our repository. You can obtain the plasmid from us or receive a ready-to-use viral preparation from the...processes. For more information, see our Guide to Plasmid Pooled Libraries . ID Name Description Genes/Insert...

...protein. With this control set, you should see your target protein IP’d in the positive control sample but not...permanently altering the structure of the protein.  Controls As with any experiment, it is critical to include...include proper positive and negative controls for an IP. When possible, perform the IP in parallel with ...interest and a sample that does not. Common negative controls include knockout cell lines or tissues that do... not in the negative control, confirming that the capture antibody is functioning as expected. In order...perform parallel IPs with beads only and an isotype control antibody. Alternative applications Oftentimes, ...In cases such as this, an epitope such as c-Myc, GFP or V5 can be used to tag the protein of interest ...

...placing FP timer DsRed-E5 under the control of Neurog3 (a gene controlling pancreatic differentiation) in ....  Table 1: Monomeric FP Timers and Associated Plasmids from Subach et al. Protein Excitation (nm) ...) Emission (nm) Brightness pKa Maturation Plasmids Fast-FT 403 (blue), 583 (red) 466 (blue), 606 (red...synthesized or long-lived transcription factors control gross changes in gene expression as new body sections...Pratik, et al. "A programmable synthetic lineage-control network that differentiates human IPSCs into glucose-sensitive...Fluorescent Protein Aggregation Catch Up On Your GFP Basics Get Advice on Choosing Your Fluorescent Protein...

... Tagging Purification Controlled Expression Reporter Plasmids Reporter Plasmids Tagging and Visualization...Visualization Purification Controlled Expression Reporter Plasmids You may also like… Plasmids 101 Addgene's Molecular...interest. Plasmid ID Promoter Inducer(s) PI Purpose Murray Lab pBEST plasmids Various Plasmids pBAD, OR2...Molecular Biology Guides Plasmids 101: E. coli strains for protein expression Plasmids 101: Common Lab E. coli...list of our bacterial CRISPR plasmids, see here . pGRG Series Various Plasmids Tn7 Transposition Nancy Craig...Series Various Plasmids T7-lacO (lactose/IPTG inducible) Flag, His, Strep, mOCR, MBP, GFP, with and without...a variety of plasmids whose expression levels have been well studied or can be controlled by small molecules...

...transforming both fusion plasmids, or mating two single haploid yeast fusion plasmid transformants into a ...in this case a reporter gene (e.g. luciferase or GFP), regardless of whether it interacts with a transcriptional...reporter system (e.g. the Gateway-compatible bait Y2H plasmid pEZY202  from Yu-Zhu Zhang’s lab) is an example...accounted for using appropriate replicates and controls (see Tips below), and by validating leads with...reporter gene activation. Including a prey-only control also provides a baseline level of reporter activation...that are known to bind serves as a good positive control to ensure that your setup works. The last thing...Yeast Expression Vectors Find Bacterial Two Hybrid Plasmids Read our Blog Post About Yeast Vectors Read our...

...pCLIPf-H2B Control Plasmid CLIP-tag Control for nuclear localization 101130 pCLIPf-Cox8A Control Plasmid CLIP-tag...pSNAPf-Cox8A Control Plasmid SNAP-tag Control for mitochondrial localization 101124 pSNAPf-H2B Control Plasmid...SNAP-tag Control for nuclear localization 101123 pSNAPf-ADRβ2 Control Plasmid SNAP-tag Control for cell...CLIP-tag Control for mitochondrial localization 101125 pCLIPf-NK1R Control Plasmid CLIP-tag Control for cell...pMCP-GPI Control Plasmid ACP/MCP-tag Control for cell surface localization 101131 pACP-GPI Control Plasmid...-tag Control for cell surface localization 101128 pACP-ADRβ2 Control Plasmid ACP/MCP-tag Control for cell...Cell-Imaging Plasmid Collection New England Biolabs Cell-Imaging Plasmid Collection Technology Plasmids Supplemental...

....org Find plant plasmids Visit our plant resource page Deposit arabidopsis plasmids with Addgene ...this method include: Cloning and transforming a plasmid into the bacterium Agrobacterium tumeficans - a...available (Weigel and Glazebrook, 2002). Find plant plasmid resources at Addgene  Choosing an expression system...options, but here are a couple of my favorites. Plasmids from the Tsuyoshi Nakagawa lab are binary gateway...terminally tagged fusion proteins with fluorescent (YFP, GFP, CFP, RFP, etc.) or affinity tags (HA, FLAG). There...prefer Golden Gate assembly, there are a series of plasmids from the Dinneny lab that additionally allow for...culture you are growing definitely contains your plasmid. Grow lots of Agrobacterium: The more agrobacterium...

...48201 CAG-GFP-IRES-CRE Cre and GFP coexpression CAG Retroviral Gage 49054 CAG-GFP/cre Cre-GFP fusion CAG...116879 CAG-Cremyc-2A-GFP GFP and Cre CAG Mammalian Lu 117148 Hiv7CMV-Cremyc-2A-GFP GFP and Cre CMV Lentiviral... Plasmid Collections Cre-lox Plasmids Cre-lox Plasmids Additional...iRFP670-EFS:Cre-2A-GFP iRFP670, Cre, and GFP TRE Mammalian Jacks 68544 AAV-Cre-GFP Cre CMV AAV Nestler...13770 pCALNL-GFP Cre dependent GFP expression Mammalian Cepko 8389 p212 pCMV-EGFP/RFP EGFP-dsRed gene switch...promoterless CRE-GFP fusion none Mammalian Sauer 11960 pBS537 tet-hCMV-GFPcre tet inducible Cre-GFP fusion tet-hCMV...shRNA expression, removal of EGFP; See also similar plasmids pSico PGK GFP and pSico PGK puro Lentiviral...

...protease, Nsp5. Our set also includes a GFP construct as a control. For easier portability, all viral genes...going on there!).  Find the Krogan Lab SARS-CoV-2 plasmids (Note: The Nsp6 construct is undergoing QC and... has been inundated with requests to share our plasmids with other researchers. We have been excited to...Gordon) have been quite busy shipping and sending plasmids to all who request them -- in locations all over...Facebook. Find more COVID-19 and coronavirus related plasmids and resources at Addgene* Many thanks to our...people, not just scientists! *If you are requesting plasmids for coronavirus or COVID-19 research, please email...

...). AAV Vectors for Serotype Testing pAAV-CAG-GFP (Plasmid #37825) Description : Ready-to-use AAV in various...These AAV are made from plasmid 37825 (deposited by Edward Boyden ) and direct GFP expression from the CAG... AAV available from Addgene's viral service. Control EGFP vectors in various serotypes for serotype testing...37825-AAVrg.T 20 µL $ 140 Add to Cart pAAV-hSyn-EGFP (Plasmid #50465) Description : Ready-to-use AAV in various...These AAV are made from plasmid 50465 (deposited by Bryan Roth ) and direct EGFP expression from the human...list of all 100 µL sized sample, please see our Control AAV Inventory . The catalog ID for the 20 µL sized... catalog item, including the titer and quality control, visit the material page (linked for each catalog...

... Investigator Plasmids GFP GFP-dependent transcription factors Connie Cepko See Plasmids Dronpa Optical...Optical control of protein activity Michael Lin See Plasmids Do you have suggestions for other plasmids that...take advantage of existing GFP-lines for cell-specific manipulation. Plasmids Fluorescent Protein Purpose...Plasmids using a fluorescent protein as a scaffold to allow scientists to selectively modulate gene expression... Plasmid Collections Fluorescent Proteins Activity Regulation Fluorescent Proteins: Activity...Optogenetics Background Fluorescent proteins such as GFP have long been used for labeling, but new methods...regulate a variety of other activities. By using GFP as a scaffold, scientists can design de novo systems...

...You’ve designed the perfect experiment – controls, conditions, and everything in between – now all you... (FLAG, Myc, etc.) and fluorescent protein tags (GFP, mCherry, etc.). Fluorescent tags are primarily used...Antibody protocols  Resources on the Addgene blog Plasmids 101: Protein tags Antibodies 101: Epitope tags...

...research? Check out Brainbow plasmids available from Addgene! Find Brainbow Plasmids @Addgene: Brainbow 1.0...1.0, 1.1, 2.1 plasmids Brainbow 3.0, 3.1, 3.2 plasmids References: Transgenic strategies for combinatorial...most important ways to spatially and temporally control gene expression, especially in in vivo models, ... of Cre-lox. Check out our previous blog post, Plasmids 101: Cre-lox, if you need a quick primer on how...-2.1 construct can express one of four colors (n-GFP, RFP, YFP or M-CFP.) The construct contains two tandem...functionally similar to Cre-based Brainbow, but is controlled by FLP/FRT recombinase. When placed under different.... The use of AAV provides spatial and temporal control without the need for germline modification, and...

...vectors in the “controls” category, which includes things like viral vectors that deliver GFP.   Figure...realized we could apply the lessons learned from our plasmid lab to develop a reliable and robust high quality...offer many resources for producing vectors from plasmids — including the protocols we use in our lab — ...while viral vectors are not as easy to ship as plasmids, they still have a global reach. From 2016–2023...vectors in our repository are produced from only 413 plasmids containing a genetic payload. While AAV9 and AAV1...having reliable ready-to-use vectors made from plasmids in the Addgene catalog would be broadly useful...conditions. Sharing our experiences Addgene’s plasmid repository gave us the experience to launch a high-quality...

...both purified CPP-tagged protein and an untagged control could enter cultured cells and activate expression...Neuronal stem cells transduced with iTOP resulting in GFP expression. Image courtesy of Diego D'Astolfo. ...10.1016/j.cell.2015.03.028. PubMed. Find the Plasmids @Addgene: iTOP-tested proteins and reporters...

...containing plasmids. Each destination vector contains GFP, enabling you to select cells with high GFP expression...multiple gRNAs on the same plasmid, you’ll make sure that each cell that gets the plasmid contains all of the...rectangles) are cloned into various plasmids using oligonucleotides. These plasmids contain Type IIS restriction...different purposes.   Gersbach Lab multiplexing plasmids: This plasmid set allows you to express 2-4 gRNAs, with...assembled to produce a plasmid encoding the gRNAs and Cas9. As with the Gersbach lab plasmids, multiple Cas9 ...Golden Gate/Gibson Assembly Multiplexing Plasmids: These plasmids allow you to assemble 2-4 gRNAs through...Golden Gate/Gibson Assembly Multiplexing Plasmids: These plasmids can be used to express up to 8 gRNAs after...

...soCoChR-GFP Syn CoChR (soma-targeted) GFP Constitutive 9 Boyden 107712 pAAV-hSynapsin-FLEX-soCoChR-GFP Syn...pAAV-hsyn-Jaws-KGC-GFP-ER2 Syn Jaws GFP Constitutive 5, 8, rg* Boyden 84445 pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] CAG...neuroscience to control electrical potentials in neurons. See our Optogenetics Plasmids and Optogenetics...* Deisseroth 58880 pAAV-Syn-ChR2(H134R)-GFP Syn ChR2/H134R GFP Constitutive 8, rg* Boyden 100054 pAAV....Constitutive 9 Harvey 59170 pAAV-Syn-Chronos-GFP Syn Chronos GFP Constitutive 1, 5, rg* Boyden 59171 pAAV-...Constitutive 8 Deisseroth 22222 AAV-FLEX-Arch-GFP CAG Arch GFP Cre dependent 1, 9 Boyden 28305 pAAV-FLEX-...dependent 5 Boyden 99039 pAAV-CamKII-ArchT-GFP (PV2527) CaMKII ArchT GFP Constitutive 1, 5, 9, rg* Boyden 137148...