... 0.72 pmol Transfer Plasmid* 1.64 pmol OptiPro SFM to total volume 500 μL *Plasmid concentrations and ... each transfer plasmid. Pro-Tip Endotoxins can inhibit transfection; therefore, plasmid DNA purification...endotoxin removal step. For high quality plasmid DNA, the plasmids should also be propagated in an endonuclease... cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would be...1: 293T cells were transfected with the GFP-expression plasmid pRosetta using μg total DNA to μg PEI ratios...channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container pH meter Stir plate...~20 h. Prepare a mixture of the 3 transfection plasmids into OptiPro SFM: Reagent Amount per 10 cm dish...

... of our control plasmids in new serotypes, as well as a new Cre dependent GFP control plasmid for expression... Cre and GFP. The GFP dependent FLPo, Flp-DOG (75469), is also available in AAV1. Plasmid Serotype .... Find the Dolcetto lentiviral prep here Control AAV Control AAV allow researchers to not only interpret...mediated neuronal control. The advantages of optogenetics lie in the spatiotemporal control offered by the...respectively of a GFP dependent Cre. The two segments come together in the presence of GFP to form a functional... pAAV-hDlx-GqDREADD-dTomato-Fishell-4 Controls Plasmid Serotype Name 114471 AAV2 pAAV-Ef1a-fDIO...mCherry 83900 AAVrg pAAV-mDlx-GFP-Fishell-1 Biosensors Plasmid Serotype Name 83899 AAVrg pAAV-mDlx-GCaMP6f-Fishell...

...GBPs/GFP. The new system, CRE-DOG (Cre Dependent On GFP), is activated by GFP and derivatives GFP and ...to bind GFP. The idea of using GFP as a scaffold suddenly seemed very realistic. Envisioning GFP as a substrate...presence of GFP. These systems, known as T-DDOG and Cre-DOG, respectively, repurpose popular GFP reporter...lines expressing GFP in specific cell types to do more than just label cells. If GFP could be co-opted...co-opted to control gene expression, he could then selectively manipulate only GFP-labeled cells. Once Tang ...tested pairs of their GFP-binding proteins (GBPs) to find those that could co-occupy GFP. Once they’d found...culture system. In cells cotransfected with GFP and the GBPs, GFP linked the two GBPs together to create a...

...scientists to begin to study GFP and deduce its biochemical properties. Now GFP and its fluorescent derivatives...biology. Scientists can easily utilize GFP-containing plasmids as a means to many functional ends. We’...fusions. Transcription Reporter: Placing GFP under the control of a promoter of interest can be used to...selection: Expression constructs like plasmids often include GFP as a marker to help identify which cells...taken up the plasmid. This can serve as an alternative to antibiotic selection. Plasmids of this type ...type may have the GFP under the control of an additional promoter from that of the gene of interest, or expressed...expressing GFP from cells that are not. Developmental/transgenic uses: Because of its stability, GFP can be...

...those formed from control (GFP gRNA-targeted) BRAF sites (lanes 3, 4) were used as controls. Duplexes were...from BRAF-edited and GFP-edited amplification products (BRAF-GFP duplex) and from GFP-edited amplification...amplification products (GFP-GFP duplex) as a control. 4. After optimizing the reaction and confirming the presence...express CAs9 and are included as a control (WT). Vinculin is a loading control. 3. In terms of making the... for gene editing. A virus encoding GFP plus a gRNA targeting GFP (pXPR_011, 59702-LV) was used to assay... by the virally-delivered GFP gRNA, will edit out the virally-delivered GFP gene. So, cells with no Cas9...I thought I could see GFP expression with a fluorescence microscope, but the GFP expression was so low...

...popular control AAV encoding GFP. Check out our inventory of serotype testing AAV New control AAVs ...about. Controls Plasmid Serotype Name 37825  AAVrg (20 µL size serotype testing) pAAV-CAG-GFP 50465... Plasmid Serotype Name 105530  AAV1 pAAV.CMV.PI.EGFP.WPRE.bGH 37825  AAV1 pAAV-CAG-GFP 59462  AAV2...Chemogenetics Plasmid Serotype Name 119472  AAV5  AV SYN PSAM4 GlyR IRES EGFP   Biosensors Plasmid Serotype...intersectional studies And more! Control AAV We’re always excited to add new control AAVs to our inventory since...optogenetics inhibitors Plasmid Serotype Name 99039  AAVrg pAAV-CamKII-ArchT-GFP (PV2527) 105679  AAV5... testing) pAAV-CAG-GFP 50465 AAV8, 9 (20 µL size serotype testing) AAV-hSyn-EGFP Biosensor AAV The...

...pAAV.hSyn-FLEX.iGABASnFR.F102G pAAV-Syn-Archon1-KGC-GFP-ER2 pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2] pAAV-FLEX-tdTomato pAAV-GFAP104...pAAV-mDlx-NLS-mRuby2 CAG-NLS-GFP pAAV-hSyn-mCherry Read on to learn more!  Calcium sensor AAV The GFP-based calcium...pAAV-Syn-Archon1-KGC-GFP-ER2) and cre-dependent version (pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2]) of this membrane-embedding...serotype you’d like to be notified about. Controls Plasmid Serotype Name 114472  AAV5, AAV8  pAAV-...indicators Plasmid Serotype Name 119036  AAV1  pAAV-hsyn-flex-Voltron-ST Chemogenetics Plasmid Serotype...37825  AAV8*, AAV9*  pAAV-CAG-GFP 50465  AAV8*, AAV9*  pAAV-hSyn-EGFP *Also coming in our new 20 ...cre-dependent version at Addgene! Fluorescent protein and control AAV These fluorescent protein expression vectors...

...the various controls that can be used for plasmid-based experiments. What are control plasmids? Generally...Negative controls: untreated cells, empty vector control, and non-targeting control Negative control conditions...follow an example of a plasmid-based experiment, delineate its requisite control plasmids, and discuss why ...Transfection controls: empty vector and internal control A transfection control measures transfection efficiency...cells. One transfection control is an empty vector control; specifically, the plasmid without the independent...empty control vector would be Plasmid A sans shRNA, or backbone Y alone. The empty vector control allows...internal control may be a plasmid that constitutively expresses a reporter protein (e.g., GFP or luciferase...

...decrease in red puncta. Find the mRFP-GFP-LC3 plasmid at Addgene! GFP-LC3-RFP-LC3∆G: A fluorescent probe ...an internal control. Autophagic flux is estimated by the GFP/RFP ratio: a decrease in GFP fluorescence...Addgene! mRFP-GFP-LC3 tandem fluorescent protein This autophagy biosensor contains mRFP and GFP fused to the...flux. mRFP-GFP-LC3 functions as an autophagy bionsensor via the same mechanism as Rosella: GFP is quenched... that releases an internal control Another autophagy biosensor is the GFP-LC3-RFP-LC3ΔG probe. It’s similar...molecule of GFP-tagged LC3 for every one molecule of RFP-tagged LC3∆G. Like the unmodified LC3, GFP-LC3 is ...decrease in autophagic flux.   Find the GFP-LC3-RFP-LC3∆G plasmid at Addgene! References Klionsky, Daniel...

...Designing the donor plasmid Figure 2: Donor plasmid design strategy. A donor plasmid contains 1 kb homology...and tested plasmids that use CRISPR/Cas9 to endogenously tag a wide variety of genes with GFP. These plasmids...found within the homology areas of the plasmid to prevent the plasmid from being cut and destroyed by CRISPR...introduced donor plasmid as a repair template. This results in the insertion of the GFP tag precisely where... the tag. Created with BioRender.com. Plasmids, plasmids everywhere To find out more about gene edited...individual proteins under endogenous regulatory control. But even with the advent of CRISPR/Cas9 technology... in human cell lines. Furthermore, the quality control necessary to ensure the edited cells are behaving...

...Chang-Deng Hu Green sfGFP Split super-folder GFP, our most-requested GFP(1-10) and GFP(11) Versatile protein...Commun. 2016 Bo Huang spGFP Split superpositive GFP Split-superpositive GFP reassembly is a fast, efficient...popular split-FP plasmids from our catalog to help you find the right split-FP plasmid for your experiment...live cells.  Controls  When designing experiments, it’s important to include proper controls to assess the...base FP (for example, EBFP2(1-10) can be used with GFP(11)). Some of the articles linked below only provide...below, and click the Article link to find all the plasmids from the associated paper:   Color Base FP...Blue EBFP2 EBFP2(1-10) and Capri(1-10) for use with GFP(11) Multiplexed labeling of cellular proteins with...

...light-based control of signaling pathways. This is particularly exciting because using light to control signaling...  Option #2 may sound like fiction, but light-controlled protein purification is now a reality thanks ..., and their use for protein purification and controlling cell signaling. Key components of OptoNBs OptoNBs...Lab monitored how AsLOV2 insertion into mCherry or GFP nanobodies affected binding of their target proteins...co-localization with a membrane-localized mCherry or GFP protein. With this system, if the OptoNB can bind...insertion into loops 1 and 6 resulted in light controlled binding, but with opposite effects: insertion...more OptoNBs, one against mCherry and one against GFP. Insertion of AsLOV2 into loop 1 always generated...

...the transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral... 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 Our 293T.... coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.   ...virus preps are titered and subjected to quality control by Addgene before being distributed to customers...production protocols, titering methods, and quality control are described below. AAV Production AAV distributed...probes targeting the ITR elements and an internal control of known titer (protocol modified from Lock et ...

...expression plasmid encoding the ORF to be expressed Finally, the researcher needs a plasmid encoding their...TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a mutant pSer-containing GFP that can be...be used as a positive control during optimization experiments. This version of GFP will contain a phosphoserine...-phosphorylated or ‘wild type’ control protein. The ‘wild type’ control can be made one of two ways. First...using the same expression plasmid. This is a particularly important control when creating a phosphoserine-incorporation...Sep-OTSλ plasmid - Addgene # 68292 C321.ΔA bacterial strain – Addgene #68306 pCRT7 NT TOPO E17TAG GFP plasmid...Find Synthetic Biology Plasmids Find Bacterial Expression Systems Find Plasmids for Protein Purification...

...released Anti-GFP [N86/8R] as another option for targeting GFP. But why might you need a second GFP antibody...western blot for mEGFP-HRas (from Plasmid 18662). See the Applications section on the Anti-GFP [N86/8R] page...of the new plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. ...  Find heat-inducible lentiviral plasmids here!   Wu Y, et. al. Control of the activity of CAR-T cells ...PMCID: PMC9015817.   Meet anti-GFP [N86/8R]! by: Ashley Waldron Anti-GFP [N86/38.1R] was among the first... missed something smokin'? You can pitch a Hot Plasmids section (antibodies and viral preps welcome!) ...system Heat-inducible lentiviral Car constructs Anti-GFP [N86/8R] ready-to-use antibody Hyperfolder YFP ...

... an inducible promoter (Plasmid 64542 or 64543) Before translation, both GFP and RFP are bound to the...Addgene.org Find Fluorescent Protein Plasmids Check Out Other Hot Plasmid Technologies ...researchers explore the complexity of translational control in cellular physiology. Ribosome profiling Ribosome...switching, TRICK requires the following components: a GFP protein that binds to the coding region of a reporter...reporter mRNA via a PP7 coat protein (NLS-PCP-GFP) an RFP protein that binds to the 3’ UTR of the reporter...appear yellow. The act of translation pushes NLS-PCP-GFP off of the mRNA, leaving only a red FP bound to the... contains a nuclear localization signal, NLS-PCP-GFP then returns to the nucleus to bind cognate untranslated...

...(cap) expressing plasmids is generated. Diversity is introduced into the cap plasmid library by inserting... comparison, capsids were packed with a GFP transgene and GFP intensity and transduction efficiency of...Caltech stepped up to the challenge. Find the PHP Plasmids Here! Their Solution: Use the CREATE method to...region of the cap gene. In this special CREATE cap plasmid, loxP sites flank the downstream polyA sequence...sequence. When viruses produced from this plasmid pool are injected into an animal with cell-type-specific Cre... do these AAVs transduce the CNS and PNS? Both plasmids infect the PNS or the CNS much better than the...AAV.PHP.eB and AAV.PHP.S had 1.5- to 2-fold higher GFP intensity compared to the strains they were derived...

...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...Science Find the Cas14 plasmids Anti-CRISPR proteins for optogenetic control of CRISPR-Cas9 Anti-CRISPR... Find the anti-CRISPR plasmids Tagging human transcription factors with GFP Kevin White’s lab recently...deposited a set of CRISPR plasmids to tag human transcription factors with GFP. As part of the Encyclopedia...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...Read the paper in Science Find the new SHERLOCK plasmids More precision with dual-nuclease Cas9-Cas9 ...Nature Communications Find the Cas9-Cas9 chimera plasmids A modular toolkit for genome editing in plants...

...recombinase. If GFP is under control of an endogenous promoter, you can use expression GFP to track cells...tag an endogenous protein with GFP, as seen in blue flame plasmid OCT4-eGFP-PGK-Puro from the Jaenisch lab...knockout a given gene and insert GFP into the genome, we’d create a plasmid similar to the one shown below...designed to insert GFP into a given locus.   Conditional knockouts The methods and plasmids described in...DNA designed in the lab. Specially constructed plasmids can be used to replace genes in yeast, mice, or...gene. Here, we'll describe the techniques and the plasmids used to inactivate specific genes in mammalian...mind when choosing a knockout strategy! Knockout plasmids Homologous recombination is a mechanism to accurately...

...dim GFPs The availability of the three dimensional structure of GFP, its brighter mutant S65T-GFP, and...and GFP shRNA. Mice carrying the R26DsRedR; CRUSH-GFP; and Nestin-Cre alleles showed efficient GFP knockdown... Hot Plasmid list. Other popular plasmids & blog posts @Addgene: The 10 Most Distributed Plasmid Technologies...inducible inactivation. Find the Robinson plasmids or the Royle plasmids and get to work! Robinson et al...editing. Dr. Masahito Ikawa has created a GFP reporter plasmid for scientists to validate the efficacy ... target sequence, the plasmid undergoes HR or SSA to reconstitute functional GFP, and the cells will turn...set of plasmids required to make singly or multiply phosphorylated proteins (B40 OTS and pCRT7-GFP). To ...