Skip to main content

We narrowed to 378 results for: proc.2

Showing: 211 - 240 of 378 results
  1. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    Published
    Oct. 24, 2017, 1:54 p.m.
    ...scientists have 1) the courage to make mistakes and 2) the integrity to recognize these mistakes quickly...She studies beta-cell function and failure in Type 2 Diabetes. Birmingham, AL, USA 33.5207°N 86.8025°W ...available to people who want to join the dialog and 2) remember that science should not be partisan: everyone...might feel disconnected from the entire scientific process.  I've spent a lot of time debating how to fix ... science in May, I marched to advocate for the process we use to learn about the world. In a way, I marched...
  2. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    Published
    July 30, 2024, 1:15 p.m.
    ...’re delivering their payload to.   Figure 2: Numbers of AAV tools, by category, available from... substrates, light, or engineered ligands (Figure 2). Biosensors, which include calcium and voltage sensors...selection process to standardize our feedback and evaluation of viral vectors. This process builds on ...significant time and removes a variable from the lengthy process of validating and using an AAV in a specific experimental...its inherent bias: Addgene uses a prioritization process to decide which tools we make available. This ...us first identifying broadly useful vectors, a process that isn’t as simple as it seems. Take, for instance... and blog posts. The lessons learned from this process have shaped and supported our growth and development...
  3. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    Published
    June 20, 2024, 1:15 p.m.
    ...the chromosome, ending replication.   Figure 2: Prokaryotic chromosomal replication. Adapted from...Microbiology and Molecular Biology Reviews: MMBR, 62(2), 434–464. https://doi.org/10.1128/MMBR.62.2.434-464.1998...control point of DNA replication in bacteria, so the process is carefully regulated by a mix of positive and...replication (oriC), where initiation begins. This process relies on the initiation protein DnaA. First, DnaA... versions is required to start the replication process. When enough copies of ATP-DnaA are bound, including...integration host factor (IHF) assists with the process: it bends the DNA to redistribute the DnaA from...plasmid ori, after which replication of the plasmid proceeds (and terminates) similarly to the bacterial chromosome...
  4. Antibodies 101: Conjugation

    Type
    Blog Post
    Published
    March 26, 2024, 1:15 p.m.
    ...residues, which can be found throughout the antibody. 2) Endogenous cysteines, such as those that make up ...Detecting antigens by quantitative immuno-PCR. Nat Protoc 2:1918–1930. https://doi.org/10.1038/nprot.2007.267 ...yourself, there are kits available to make the process quick and easy. Before getting started, be sure...
  5. BeHeard Award 2018: Diseases of Glycosylation, Arginine Mutagenesis, & Neural Development

    Type
    Blog Post
    Published
    June 26, 2018, 1:33 p.m.
    ...studies have shown that the conditions Sotos-type 2 and Marshall Smith Syndrome are due to mutations in...accurate diagnosis of NFIX-related syndromes (Sotos-type 2 and Marshall-Smith) and could be expanded to NFIA-...lines and use these lines to study how immune processes and biological pathways are altered in PMM2-CDG...
  6. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    Published
    Sept. 17, 2024, 1:15 p.m.
    ... that all look exactly alike.      Figure 2: Reagent gradient. Here, the lanes are lightly marked...guide. The gel is then removed from the membrane (2) and the membrane is cut so each lane is its own piece...in the bands, you may want to consult Tables 1 and 2 in our Technical Design of a Western Blot post to ...the antibody’s dynamic range. Specifically, the 1:2, 1:1, and No dilution samples all have approximately...conditions are 4 °C overnight or room temperature for 1–2 hours, there is some evidence that antibodies may ...high background or low signal during the imaging process, try adjusting your exposure time first, since ...
  7. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    Published
    Oct. 10, 2023, 1 p.m.
    ...antibiotic selection. Essentially, this gene is a 2-in-1 providing both bacterial and eukaryotic selection...Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi.org/10.1093/mutage/gei016I .... During the propagation and/or transformation process, some bacteria will either never receive a copy...antibiotics listed below affect continuous cellular processes (e.g., protein synthesis), while amp only affects...
  8. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    Published
    Dec. 7, 2021, 2:15 p.m.
    ...green squares, purple circles and blue triangles. 2) An antibody immobilized on a bead binds specifically...protein G, a novel IgG-binding reagent. J Immunol 133(2):969-974. Hjelm H, Hjelm K, Sjöquist J (1972) Protein...to immunoprecipitation and simply refers to the process of anchoring an antibody, often called the capture...protein interactions, and limit high throughput procedures. However, agarose beads are often a more cost-effective...
  9. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ...with neuron-specific expression of dCas9-VPR (Figure 2), a popular Cas9 activator developed by the Church...ubiquitous ones such as EF-1α and CAG. Figure 2: Dual lentivirus system with ubiquitous expression..., studying the genetic underpinnings of these processes has been complicated by the lack of precise genetic...undifferentiated cells at the end of the screening process. However, while the neuronal marker TUJ1 is useful...
  10. Deaminet 2026: Breakthroughs in Base Editing, Deaminase Biology, and Therapeutic Translation

    Type
    Blog Post
    Published
    March 10, 2026, 1:14 p.m.
    ... a new generation of deaminases. Figure 2: Structure of ABE8e*A showing dimerized TadA subunits...et al. (2024) under a CC BY-NC 4.0 License.  2. Precision and safety: A new generation of off-target...191–202. https://doi.org/10.1038/s41586-025-09732-2 Richter, M. F., Zhao, K. T., Eton, E., Lapinaite, ...resolution, to cancer biologists dissecting mutational processes, to medical researchers advancing base and prime...Understanding deaminases at atomic resolution PE6d: Processivity gains and overextension trade-offs The prime...editor PE6d exhibits substantially increased processivity and editing efficiency relative to PE2 (Doman...alone, indicating an intrinsic shift in enzyme processivity. Structural data implicate specific residues...
  11. Antibodies 101: Choosing the Right Isotype

    Type
    Blog Post
    Published
    April 6, 2023, 3 p.m.
    ... reduce the background staining (Table 2).      Table 2: Secondary antibody selections for a four-antibody...have been cross-adsorbed. The cross-adsorption process filters out off-target antibodies increasing specificity...
  12. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ...wheat germ, HeLa, and Leishmania (Gagoski, et al 2015[2]). The proteins encoded in this set range in size ...to generate Illumina sequencing libraries using a 2-step PCR-based protocol and are compatible with single-end...decrease false positives observed in the cloning process, making it easier to produce the constructs you...novel and powerful pyranopyridine derivatives." Proceedings of the National Academy of Sciences 113.13 (2016...-time observation of metabolite production." Proceedings of the National Academy of Sciences 113.9 (2016...pCri system to simplify the protein purification process by making it easy for a researcher to quickly generate...on the system Easing the Protein Purification Process with pCri Cell-free protein expression kit Cell-free...
  13. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ...break), increasing editing efficiencies 2-3-fold. However, the process of double nicking increases indel formation... editing as efficient as possible. Figure 2: Decision tree for selecting a PE6 variant. PE = prime...using paired prime editing. Nature Biotechnology, 40(2), 218–226. https://doi.org/10.1038/s41587-021-01025...the template and primer binding site, increase processivity, or improve thermostability. What came out on...repair (MMR) complex, to temporarily inhibit this process and promote copying of the edited strand to the...to the numerous innovations that streamline the process of prime editing, the Liu lab contributed an improved...
  14. The Addgene Antibody Data Hub

    Type
    Blog Post
    Published
    Sept. 12, 2024, 1:15 p.m.
    ...whether the results include knockout data.    Figure 2: Search options in the Antibody Data Hub. Data reports...antibody worked in your application. The submission process is easy, with simple, descriptive fields, drop-..., and can be saved at any point throughout the process, so you can work through it at your own pace. To...
  15. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    Published
    Oct. 6, 2020, 1:15 p.m.
    ...gene activation, with some genes experiencing around 2-fold activation levels. Although other methods have.... This method has been shown to activate genes at 2- to 3-fold higher than SAM, SunTag, and VPR (Zhou ...activation across multiple loci was an arduous process. When using zinc finger proteins or TALE proteins...
  16. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    Published
    Aug. 11, 2016, 2:23 p.m.
    ... digest and compare it to some reliable controls. 2. Viral stocks may be sensitive to freeze thaw cycles...PMID: 24003179. PubMed Central PMCID: PMC3760253. 2. Rohwer, Forest. "Global phage diversity." Cell 113.2...volume. Always remove packaging cell debris before proceeding to ultracentrifugation to avoid contaminating...
  17. Pooled Library Amplifications

    Type
    Blog Post
    Published
    May 30, 2024, 1:30 p.m.
    ...estimate transformation efficiency (Figure 2).    Figure 2: Example of a serial dilution to estimate... reduce recombination during the amplification process. In liquid culture, all of the bacteria can grow...
  18. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ... PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an insertion locus-specific gRNA The gRNA ...Find the plasmids from this publication at Addgene. 2. Dalvai, Mathieu, et al. (2015). “A Scalable Genome-Editing...MMEJ doesn’t get a lot of press. However, this process accounts for a percentage of the mutations seen...
  19. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    Published
    May 15, 2014, 3:33 p.m.
    ...from GFP and the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising..., V163A EBFP F64L, S65T, Y66H, Y145F   Table 2: Functional role of specific mutations in GFP derivatives... enabling the study of dynamic, physiological processes. Re-engineering GFP to increase its range of color...
  20. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    Published
    Nov. 3, 2016, 2:30 p.m.
    ...Information Science and Technology 60.4 (2009): 858-862. 2. Van Noorden, Richard. "Interdisciplinary research...Librarianship and Scholarly Communication 2.2 (2014): 2. Additional Resources on the Addgene Blog 6 Tools...others for failing to find our work?  I study processes for diagnosing and resolving organizational performance...impact of their contribution. It offers a 5-step process and several examples for creating effective research...
  21. March for Science

    Type
    Blog Post
    Published
    April 21, 2017, 2:30 p.m.
    ...citizens enter into in order to protect their rights (2). Less formally, governments are the organizations...science education and literacy that this changes.  2) Lumping all scientists (and consequently their results...politics? Science is an apolitical process for seeking knowledge. The process begins with a testable hypothesis...hypotheses and the cycle begins again. No part of this process is inherently political so why is there a march...scientists as well as other citizens. Thus civilization proceeds in an informed fashion. For science to be useful...
  22. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    Published
    Nov. 3, 2022, 12:15 p.m.
    ...faster when using a dual guide system.   Fig. 2: Size screening for large deletion events.   ... Biol 17: https://doi.org/10.1186/s13059-016-1012-2 Find more CRISPR Resources at Addgene: Read our CRISPRGuide...looking for, you are ready to begin the screening process. Want to set yourself up for a screening success...then a single, bulk validation may be enough to proceed with your experimental plan. Validating knockout...
  23. Countdown to Halloween @Addgene

    Type
    Blog Post
    Published
    Oct. 29, 2014, 5:26 p.m.
    ...Halloween is only 2 days away and here at Addgene we couldn’t be more excited! As a small, tight-knit...suggestions for this year’s teams that may be procrastinating choosing their costume?  As our organization...
  24. Let There Be LITE Plasmids

    Type
    Blog Post
    Published
    Nov. 21, 2013, 3:56 p.m.
    ...First, the light-responsive molecules of cryptochrome 2 are the chromophores flavin-adenine dinucleotide and...of optogenetic control of endogenous cellular processes." The LITE system can now be applied to unraveling...genetic and epigenetic players in normal biological processes and disease states. While LITE plasmids have clear...
  25. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    Published
    June 27, 2024, 1:15 p.m.
    ...would label everything but the CD8+ T cells (Figure 2). Even better, we can use the same fluorophore for...gating on the negative population.     Figure 2: Dump gating labels all unwanted cell lineages with... for entry of the antibodies. Here, a two-step process is employed: the cells are first fixed and then...phosphatases, and ubiquitin ligases; and many more. This process, however, leaves the nucleus intact. If markers...not need to use a separate fix/perm cytosolic procedure in that case.     Figure 3: For intracellular...
  26. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    Published
    Sept. 28, 2023, 1:15 p.m.
    ...to a specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID ...cells will be ready for the spotlight.    Figure 2: Opto-RhoGEFs to control Rho GTPase activity. A) Schematic...great way to stimulate and study these physical processes. Joachim Goedhart and colleagues in the Gadella...could be applied to study other actin-dependent processes in cells and tissues. And, this Opto-RhoGEF design...the protein of interest supports a variety of processes in different cell types. One great way to do this...
  27. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 4, 2017, 2:30 p.m.
    ...linear region to solve for total expression time. 2. Timer functionality with a single gene Although it...290.5496 (2000): 1585-1588. PubMed PMID: 11090358. 2. Tsuboi, Takashi, et al. "Age-dependent preferential...allowing them to optimize the compound production process. How have you used FP timers? Are you planning ...
  28. Getting to Know Expansion Microscopy

    Type
    Blog Post
    Published
    June 20, 2023, 1:15 p.m.
    ...your sample (i.e. cultured cells or tissues).  Step 2: attach anchors to your sample that will link biomolecules...quite so much. Created with biorender.com Steps 2-4 are not found in standard IHC/ICC protocols and ....)  Step 5: Image   Figure 1: The ExM process allows you to physically magnify a sample so you...
  29. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    Published
    Sept. 17, 2019, 12:40 p.m.
    ...of these neurons via the channel Channelrhodopsin 2 (ChR2) reduced glutamate transmission from the mPFC...deliver Cre-dependent expression of  Channelrhodopsin 2 (ChR2), a modified receptor protein that activates...tracking software. Locomotion is a highly complex process on the level of individual neurons and in terms...
  30. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    Published
    March 8, 2021, 2:41 p.m.
    ... a western, proteins are: (1) separated by size, (2) transferred to a membrane, and  (3) detected using...expensive system-specific sandwiches.   Figure 2: The western blot dry apparatus includes a stack of...gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences...  Figure 1: Overview of the Western blot process. The sample, a mixture of protein is separated ...antibodies, and finally visualized.   This process allows you to detect a single, specific protein...conduct the electric current. The wet transfer process is very effective in transferring a broad range...common readout for subcellular fractionations, a procedure that isolates proteins from different cellular...
Showing: 211 - 240 of 378 results