We narrowed to 584 results for: abo.1
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TypeBlog PostPublishedJuly 16, 2020, 1:15 p.m....was around 33% of that observed between Jan. 1 and Mar. 1, 2020. Figure 2: A simple approximation...paper. Green and red lines depict the sum of mean +1 and -1 std for each paper, respectively. The fitted...complication in the China data – there is a period of 1 month, from 1/22/20–2/23/20, where there were zero plasmid... from 5/22/14 to 5/19/20). Each paper has between 1–32 plasmids and a total of 2–2,590 requests. We received...correspond to publications of popular papers. Figure 1: Plasmid requests by date. Parentheses depict the ...China. However, it is evident that if we believe that 1/22/20–2/23/20 are the more accurate dates of lockdown...in his laboratory. Learn more here. Additional resources on the Addgene blog Learn more about plasmid...
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MXS Chaining
TypeBlog PostPublishedFeb. 7, 2017, 3:30 p.m....assembly. Journal of biological engineering. 2010;4(1):1. Pubmed PMID: 20205762. PubMed Central PMCID: PMC2822740...with minimal spectral overlap between them (Table 1). In each construct, 3 copies of the fluorescent protein... enabled labeling of particular structures (Table 1). Each construct was flanked with a CMV promoter (...construct was then introduced to HeLa cells (Figure 1). The resulting HeLa cells show robust labeling, with...fluorophore at the respective subcellular structure (Figure 1). Here, MXS Chaining gave researchers the ability ... all four constructs into one 15 kb insert. Table 1: MXS Constructs for Labeling Cellular Structures ...maxima Tethering partner Subcellular localization 1 TagBFP 399nm/ 456nm histone 2B (H2B) Chromatin 2... -
Recombinase-based State Machines Enable Order-dependent Logic in vivo
TypeBlog PostPublishedJuly 28, 2016, 2:30 p.m....recombinase-based state machines in living cells” (1), into a more straight-forward, concise explanation...an RSM is to sequence the register. In our paper (1), we allowed individual bacterial cells to form colonies...in the Materials and Methods section of our paper (1). Lastly, it is possible to build gene regulatory ... not. How might one implement RSMs? In our study (1), we implement RSMs in E. coli with two plasmids (...inputs to endogenous gene expression (2). Our paper (1) explains how to arrange attB-attP pairs on a register...performance data from the indicated figures in our paper (1)) Input Plasmid Output Plasmid Fig. 3A pNR64 pNR160...in the Material and methods section of our paper (1). How might one troubleshoot GSRMs that don't work... -
Deep Mutational Scanning with One Pot Saturation Mutagenesis
TypeBlog PostPublishedFeb. 22, 2017, 3:30 p.m....introduce all desired codon changes (step 1 in the image above). A second universal primer is used to synthesize...strand. Both produced about the same number of positive clones. See Supplementary Note 1 in Wrenbeck et al...strand of DNA degraded in the first digest (step 1 in above diagram). Remember that the BbvCI enzymes nick...think about one pot saturation mutagenesis? How would you use it in your research? References 1. Natarajan...making some or all of the following modifications: 1) increased concentration of Taq polymerase, 2) increased...introduced, and scalability (see Supplementary Table 1 of Wrenbeck et al). It uses a uracil labeled wild-...strand is nicked by the BbvCI variant not used in step 1. Then this strand is degraded with ExoIII and ExoI... -
When Fidelity Matters: A frank discussion about ligase fidelity
TypeBlog PostPublishedMarch 6, 2018, 1:49 p.m....phosphorylated DNA termini to 3 ́-OH DNA termini (1-4). In vitro, ligases (notably T4 DNA ligase) are ... enzymology of DNA repair enzymes. References 1. Shuman, Stewart, and Christopher D. Lima. "The polynucleotide...thermostable DNA ligase-encoding gene." Gene 109.1 (1991): 1-11. PubMed PMID: 1756968. 7. Lauer, G., et al. "Cloning... And what is ligase fidelity anyway? Let’s talk about it. DNA ligases are enzymes that seal breaks in ...to accelerate characterization of nucleic acid metabolic enzymes." Nucleic acids research 44.2 (2015): ... -
The Breast Cancer Microenvironment: A Tumor’s Backstage Team
TypeBlog PostPublishedJan. 31, 2019, 1:32 p.m....enzyme (among others): caveolin 1, also known as CAV 1. Usually, caveolin 1 promotes the cell life cycle ...caveolin 1, there’s less malignancy because there’s less proliferation. However, the lack of caveolin 1 in ...believes that miRNA may decrease the expression of TAK 1, an enzyme that controls apoptosis and regulates transcription...Agnieszka K., et al. "An absence of stromal caveolin-1 expression predicts early tumor recurrence and poor... related blog posts Learn about our cancer collection pages Read more about the tumor microenvironment...the tumor. How do microenvironment enzymes and metabolic reprograming influence tumors development? Being... vice versa, let’s take a look at cancer cell metabolism. Cancer cells promote oxidative stress leading... -
Important Considerations When Using AAVs
TypeBlog PostPublishedJune 13, 2017, 2:30 p.m....in movement. Follow her on twitter @katrinaboards. References 1. Russell, David W. "AAV vectors, insertional...vectors without specific facilities (Biosafety Level-1). Other possible tools, lentiviruses for example, ...suspected cases of insertional mutagenesis in humans (1) and AAVs are non-enveloped viruses, so they are resistant...Subscribe to Viral Vector Blog Posts from Addgene 1. Location, location, location! What do buying real...important factors related to location. Location (1): What tissue? Location (2): Where in the tissue/which...the right serotype? Keep asking questions! Table 1: AAV Serotypes and Tropism Experiment Detection...the vicinity of the injection. Examining serotypes 1, 8 and 9, not only do they undergo retrograde and ... -
Live and Let Dye: Self-Labeling Protein Tags
TypeBlog PostPublishedApril 16, 2024, 1:15 p.m....intracellular targets. Nature Communications, 7(1), Article 1. https://doi.org/10.1038/ncomms10778 Erdmann...molecules in vivo. Nature Biotechnology, 21(1), Article 1. https://doi.org/10.1038/nbt765 Lavis, L. (2016... envelope design. Nature Communications, 14(1), Article 1. https://doi.org/10.1038/s41467-023-40788-8 ... capping protein. Nature Communications, 14(1), Article 1. https://doi.org/10.1038/s41467-023-39655-3 ...up linked to whatever molecule you choose (Figure 1). Just express the fusion protein and add a specific...proteins, opening up new applications. Figure 1: Live-cell labeling strategies. The green structure...minimally-disruptive genetic tag on the protein of interest. Table 1: Summary of popular self-labeling tag systems ... -
Lentiviral Vector Uses and Overview
TypeBlog PostPublishedMay 19, 2016, 2:30 p.m....HIV-1 were transduced with a lentiviral vector containing an antisense sequence against the HIV-1 envelope...Addgene depositor Didier Trono and collaborators turned to the lentivirus HIV-1, well-known to infect non-dividing...plasmids. The packaging plasmid carried a mutated HIV-1 provirus that couldn’t package itself due to a few...plasmid encoded the desired transgene flanked by HIV-1 long terminal repeats (LTRs) that facilitate viral... targeting and tracing cells in the brain (Figure 1). 2nd generation lentiviral packaging system The graphic...infectivity, is often used as a substitute for HIV-1 Env, which can only infect CD4+ cells. 3rd generation...lentivirus in the comments section below! References 1. In vivo gene delivery and stable transduction of ... -
Viral Vectors 101: Producing Your rAAV
TypeBlog PostPublishedJuly 16, 2024, 1:15 p.m....Inactivation of AAV Serotype 1 Vectors. Human Gene Therapy Methods, 28(1), Article 1. https://doi.org/10.1089...called “triple plasmid transfection” (Figure 1). Figure 1: Overview of key steps of AAV production, ...repeat sequences. Human Gene Therapy Methods, 23(1), Article 1. https://doi.org/10.1089/hgtb.2011.034 Benyamini...in the CNS. Molecular Neurodegeneration, 15(1), Article 1. https://doi.org/10.1186/s13024-020-00361-z ...dehydration (Howard & Harvey, 2017). You can learn more about common AAV applications by reading our Viral Vectors... -
Mouse Modeling, Part 2: Breeding and Crossing Mice
TypeBlog PostPublishedAug. 6, 2019, 12:55 p.m....approximately 1 in 16 mice that harbor homozygous alleles at both of your genes of interest. About 1 in 4 mice...In Part 1 of our mouse modeling blog series, we covered techniques that can be used to introduce genetic...mice in your experiment. As you’ve learned in Part 1, there are many types of genetically engineered mice..., and assuming Mendalian patterns of inheritance: 1. Genotype your parental mice In order to ensure that...example of mice that are derived from crosses. In Part 1, we explained how genetic recombination in mice expressing...expressed, or inducible (this is explained more in Part 1 of this series). In any case, Cre is a dominant phenotype...Find our Plasmids 101 blog posts Read blog posts about using Cre-lox Read our genome engineering blog posts... -
When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells
TypeBlog PostPublishedApril 19, 2016, 2:30 p.m....Figure 1). Figure 1: Each problem with dimerizing fluorescent proteins is illustrated. 1) Membrane... in nature are prone to dimerization (i.e. EGFP) [1, 2] or even form obligate tetramers (i.e. DsRed) [...active state due to closure by FP dimerization. 1. Transmembrane fluorescent protein fusions Transmembrane...bright, non-physiological, and difficult to ignore [1, 2]. 2. Fusion to an obligate dimer or oligomer Several... For a more detailed discussion see [5]. Problems 1 and 2 depend on protein concentrations. An imperfect... distance runner, gardner, and cook. References 1. Snapp, E.L., et al., Formation of stacked ER cisternae...sold by Clontech and is in a freezer box in just about every lab in the world, is not inert. In all seriousness... -
Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer
TypeBlog PostPublishedJuly 6, 2017, 1:59 p.m....aggregation, and potentiates the effect of PD-1 blockade.“ (Ref 4). PD-1 helps tumor cells evade the immune system...addgene.org to find out more about our free deposit and distribution service! References: 1. Pietrocola, Federico...fasting has a positive effect on cancer treatment (Ref 1). As described in the presentation, starvation or ...of the immunosuppressive cell surface receptor PD-1. This immunosuppressive environment in turn may represent... (Chimeric Antigen Receptors) CAR T cells and PD-1 modulators. An exciting development in immunotherapy...Cancer, Inflammation and Immunity and learn more about “... recent findings in basic immunology, cancer-immune...curing cancer in cross-disciplinary projects and collaborations. Summarized by Professor Michael Karin from... -
CRISPR-mediated Plant Base Editors
TypeBlog PostPublishedJan. 3, 2019, 1:35 p.m....rice crop improvement in the laboratory of Yinong Yang at PSU. References 1. Komor, Alexis C., et al. ...excision repair mediated by uracil glycosylase (Figure 1). The presence of a uracil glycosylase inhibitor (...generation of the desired T-A base pair. Figure 1: Transition mutation (C>T, G>A, A>G, and T>C) installation... A-T base pair is converted to a G-C pair (Figure 1). As the cellular inosine excision repair is not as...Target-AID based (CBE-2)- and rAPOBEC based (CBE-1)- CBEs are successful base editors in rice, tomato...deaminase can edit in a stretched window ranging from 1-17 bp relative to the PAM sequence (Zong et al., 2018...which recognize PAM sequences other than NGG (Table-1). Type Nickase Cas9 (D10A) Deaminase PAM requirements... -
An Integrin Antibody Toolkit from IPI
TypeBlog PostPublishedMarch 28, 2024, 1:15 p.m....-Integrin alpha 5 beta 1 [IPI-ITGA5/ITGB1.2] Integrin alpha-5,Integrin beta-1 Human, Mouse IgG1 Rabbit...-Integrin alpha 5 beta 1 [IPI-ITGA5/ITGB1.4] Integrin alpha-5,Integrin beta-1 Human, Mouse IgG1 Rabbit...specific integrins in cellular processes. Figure 1: Integrins can shape into three conformations: (A)...immunoprecipitation and immunohistochemistry. Table 1: IPI antibodies Antibody Target Proteins Reactivity...Human IgG1 Rabbit Anti-Integrin alpha M [IPI-LM2/1] Integrin alpha-M Human IgG1 Rabbit Anti-Integrin...Human IgG2a Mouse Anti-Integrin alpha M [IPI-LM2/1] Integrin alpha-M Human IgG2a Mouse Anti-Integrin... β1 integrin mechanics. Nature Communications, 13(1), 7471. https://doi.org/10.1038/s41467-022-35173-w... -
Antibody Validation for Flow Cytometry
TypeBlog PostPublishedSept. 10, 2024, 1:15 p.m....dye, respectively. WT and KO cells were mixed in a 1:1 ratio and fixed in 4% PFA and permeabilized in 0.1%...antibodies were diluted to 1 µg/mL except for 14558, which was used at 0.35 µg/mL (1/100). * = monoclonal antibody...through their HLDA workshops. Let’s use TIM-1 (Single pass type-1 membrane protein), which has been designated...specific antibody:target combination of interest (Figure 1). It can therefore be helpful to try all three when...to label an intracellular target. Figure 1: HCT 116 WT and SYT1 KO cells were labelled with a...cells were stained with the indicated Synaptotagmin-1 antibodies and corresponding Multi-rAb CoraLite® Plus...between the wild-type and the KO population (Figure 1), further optimization can be performed to both maximize... -
CRISPR-based Genome Editing Systems For Editing in E. coli
TypeBlog PostPublishedApril 13, 2021, 1:15 p.m.... Then plasmids are cured from the cell. Steps 1-1 and 1-2 can be done simultaneously. Image from Li et...coli K-12 strains, 2 E. coli B strains, 1 E. coli W strain, and 1 Tatumella citrea, with a total of 70 sites... update the pCas/pTargetF system. Figure 1: The editing process begins with the addition of the...updates involved the following changes. In pCas9, we 1) replaced the promoter of gRNA-pMB1 with PrhaB, 2)...biosynthesis, we constructed a strain library with 1-11 copies of integrated genes. We obtained a stable...complete editing process. The MUCICAT system While the above systems are great for experiments that require only...serve to interrupt genes (or gene clusters) in metabolic engineering. So far, we’ve used the MUCICAT system... -
Supporting Rare Disease Research with BeHeard 2017
TypeBlog PostPublishedJuly 20, 2017, 2:30 p.m....estimated to affect anywhere from 1 in 6,000 to 1 in 19,000 live births (1). While disruption of the smn1... funding opportunities. References 1. Jones, C., et al. "PP09. 1–2352: Systematic review of incidence...We often hear about breakthroughs in diabetes and lymphoma research or the many different ways you can...little-known disease. You can find more information about the disease on the Meothelioma Cancer Alliance Website... -
Antibodies 101: Designing Your First Flow Panel
TypeBlog PostPublishedMay 28, 2024, 1:45 p.m....marker. Looking at Table 1, we can recognize a couple of fluorophores from Figure 1. For example, the chart...panel, we'll talk about one last principle of color emission. As you can see in Figure 1, emission spectra...efficient (i.e., happen most of the time). Figure 1 shows a graph from the BD® Spectrum Viewer, outlining...Resonance Energy Transfer (FRET). Figure 1: Emission spectra and respective colors of a few commonly... the brightness of individual fluorophores. Table 1 describes the brightness of a few commonly used fluorophores...Brighter fluorophores have a stronger signal. Table 1: Degrees of brightness for a few commonly used fluorophores...blue laser excites at a wavelength of 488 nm (Table 1), which is near a local maximum of PE's excitation... -
Addgene is Expanding Our Viral Vector Service!
TypeBlog PostPublishedOct. 15, 2024, 1:15 p.m.... Request review: 1–2 days Prep and QC completed: 6–8 weeks Average shipping time: 1–2 days (USA); 3–4...choice of five different serotypes. Figure 1: The process of requesting and ordering a Packaged...request in three different quantities: 10x 100 µL (1 mL), 25x 100 µL (2.5 mL), or 50x 100 µL (5 mL). Once...with our production process. This process will take 1–2 days, and we anticipate approving the vast majority...educational resources. We hope you’re as excited about this expansion as we are! Happy packaging! ...