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We narrowed to 401 results for: ARA-2

Showing: 201 - 240 of 401 results
  1. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ... Zetsche et al., Nat Biotechnol. 2015 Feb 2;33(2):139-42. doi: 10.1038/nbt.3149. Light-inducible ...and on your favourite signalling pathways (Figure 2). They are all available now at Addgene. 1 Lungu...20;19(4):507-17. 10.1016/j.chembiol.2012.02.006. 2 Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; ...Nihongaki et al., Chemistry and Biology. 2015 Feb 19; 22(2):169-74. High efficiency cloning for enzyme screening...inducible, gene knockdown with shRNA or shRNA-miR30, (2) tamoxifen-inducible CreERT2 gene deletion, (3) simultaneous...different fluorescent proteins, 8 constitutive promoters, 2 includible promoters, 3 polyA terminators and various...of plasmids required for each input/output system; 2) removing all chemically inducible promoters; 3) improving...
  2. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

    Type
    Blog Post
    Published
    July 6, 2017, 1:59 p.m.
    ...represent a favorable state for tumor progression (Ref 2). Future treatments may need to target this immunosuppressive...cell 30.1 (2016): 147-160. PubMed PMID: 27411589. 2. Todoric, Jelena, Laura Antonucci, and Michael Karin...the Relationship” by our guest blogger Subhadra Jayaraman, a doctoral candidate at Binghamton University...
  3. Antibodies 101: Flow Cytometry Controls

    Type
    Blog Post
    Published
    June 6, 2024, 1:50 p.m.
    ...populations that may otherwise be difficult to separate out. In Figure 2, you can see that, without the FMO, it... from the Ly6C positive population. Figure 2: An FMO gating example. The FMO sample (A) can be ...
  4. CRISPR 101: Off-Target Effects

    Type
    Blog Post
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    Feb. 8, 2024, 2:15 p.m.
    ...knock-out cell line to perform downstream characterization of, isolating 2 or 3 distinct clones to perform key...sites would confound your experimental results and 2) perform targeted off-target measurements at those...experiments where only localization of the Cas occurs. 2 gRNA approach Off-targets that come from mutations...
  5. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ...somewhere else (Fig. 2). Class II TEs have LTRs on both of their ends.    Figure 2: Overview of DNA...has four major components: 1) a CRISPR RNA (crRNA), 2) four proteins (TniQ, Cas8, Cas7, Cas6) that forms...another. This jumping is what caught the eye of Barabara McClinntock who first discovered transposons due...
  6. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    Published
    Aug. 13, 2024, 1:15 p.m.
    ...detection, and control. Int J Pharm. 2018 Oct 25;550(1-2):251-268. doi: 10.1016/j.ijpharm.2018.08.043. Epub...and particle formation. J Pharm Sci. 2015 Feb;104(2):602-11. doi: 10.1002/jps.24259. Epub 2014 Nov 21....Roberts, Prasad S. Sarangapani, Protein aggregation, particle formation, characterization & rheology, Current...are contained to one plasmid or if they’ve been separated onto two. You’ll first transfect your cells with...harvesting recombinant antibodies consists of separating antibodies from media, cells, and debris. Affinity...bind ligands on the beads and a magnet is used to separate the beads from the media (Brechmann et al. al.... of your prep. In this method, antibodies are separated by size on an SDS-PAGE gel. Once stained, the ...
  7. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    Published
    Dec. 14, 2021, 2:15 p.m.
    ...Hrit et al. 2018, Kaas et al. 2013). Figure 2: CRISPR TET-based epigenetic editing. In reality, ...approach for targeted DNA demethylation. Cell Discov 2, 16009. https://doi.org/10.1038/celldisc.2016.9.  ...hear it (whether at a scientific conference or a karaoke bar), you probably want to sigh. The problem is... directly inside the gRNA sequence is a great guarantee that DNMT1 will be blocked (within PAM works, ...
  8. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    Published
    Feb. 4, 2020, 2:15 p.m.
    ...amilCP, tsPurple, and gfasPurple) (Figure 2). Figure 2: Chromoprotein expression from bacteria on.... https://doi.org/10.1007/bf00428249 Chan MCY, Karasawa S, Mizuno H, Bosanac I, Ho D, Privé GG, Miyawaki...Miyawaki A, Ikura M (2006) Structural Characterization of a Blue Chromoprotein and Its Yellow Mutant from ...Hoegh-Guldberg O (1995) Isolation and Partial Characterization of the Pink and Blue Pigments of Pocilloporid...
  9. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    Published
    Jan. 30, 2024, 2:15 p.m.
    ...Indicators GEVIs can be divided up structurally into 2 main groups: voltage sensitive domains and Rhodopsin...depending on your experimental system.   Figure 2: General overview of chemigenetic tools. A genetically...mechanism of their voltage sensing component, which separates them into two distinct categories detailed below...phosphatases from sea squirts. This design structurally separates the two components of the indicator, which can...FP-rhodopsin GEVIs have been engineered to have kinetics comparable to its one component counterpart and is significantly...
  10. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    Published
    Aug. 30, 2022, 1:15 p.m.
    ... of permeabilization and membrane integrity (Fig. 2). For example, saponin interacts with cholesterol ... at too high a concentration.      Figure 2: Examples of different permeabilization strategies...cause some confusion. Paraformaldehyde: Many IF protocols call for paraformaldehyde (PFA) as the fixative...recover good antibody binding by tweaking those parameters, but in some cases your antibody may just not...
  11. 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... color also exist with only slight variations separating them.  Please note that many fluorescent proteins...(FACS): This is a type of flow cytometry that separates mixtures of cells into distinct populations based... fluorescent signal. Thus, FACS can be used to separate cells expressing GFP from cells that are not. ...
  12. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    Published
    April 14, 2016, 2:30 p.m.
    ...be decorated with proteins and remain functional, 2) break membranes outside of their natural context ...PubMed PMID: 3759909. PubMed Central PMCID: PMC213417. 2. Schrallhammer, M. et al. Tracing the role of R-bodies...toxicity of R-body producing recombinant E. coli on paramecia. European Journal of Protistology 48, 290–296 ...
  13. 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 ...to ~4-10x linearly! This expansion physically separates molecules while maintaining their relative positions...pre-expanded samples can sometimes take on a fragmented appearance post-expansion, but that doesn’t mean we should...understanding of neuronal synapses, centrosomes, parasites, and more (Bertiaux, et al. 2021; Gambarotto, ...new insights into the cytoskeleton of malaria parasites including the conservation of a conoid. PLoS Biology...10.1038/s41592-018-0238-1  Klimas A, Gallagher BR, Wijesekara P, et al (2023) Magnify is a universal molecular...
  14. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...domains retain their individual activities when separated (2), and that DBD and TAs from different systems...308. PubMed PMID: 6312261. 2. Keegan, L., Gill, G., and Ptashne, M. Separation of DNA binding from the transcription-activating...PubMed PMID: 2547163. 5. Ozawa, T., Kaihara, A., Sato, M., Tachihara, K., Umezawa, Y.l. Split luciferase...possibility for problems with one or more of these parameters can lead to a high number of false negatives ...
  15. Advice for Starting a Biotech Company from a Grad Student turned Entrepreneur

    Type
    Blog Post
    Published
    July 10, 2014, 7:17 p.m.
    ...go anywhere. The team should initially consist of 2-3 individuals that are aligned in vision and outcomes... the technology. This transition takes around 1 – 2 years. The product is the measure of success. Technology...constantly practicing. “The expenditure of effort in preparation and practice is directly related to success presenting...
  16. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    Published
    Feb. 18, 2014, 4:22 p.m.
    ...The groups in these programs typically consist of 1-2 mentors and 3-5 mentees who meet together for an 8... together. If you all work at the same place, a 1-2 hour lunch on a regular schedule might work. If not...pre-reads, videos or exercises to be completed in preparation for the meeting or to be read or watched during...
  17. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    Published
    July 14, 2021, 1:15 p.m.
    ...animal is given additional booster immunizations every 2-3 weeks to increase the antibody titer. The titer ...level of antibodies is reached - typically within 2-4 months. After the animal has the desired level of...animal. The red blood cells and the serum are separated, and the serum - which contains the antibodies...
  18. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    Published
    Dec. 7, 2020, 2:53 p.m.
    ...et al., 2020). Genomic targets require an upstream 2-bp PAM sequence for recognition by QCascade; although...cells of the plasmid after integration. Figure 2: (A) Streamlined VchINT expression constructs. (B)...target DNA sequences using RNA-DNA base pairing. Separately, TnsABC forms a transposase complex with the ...bp target-site duplication (TSD), a hallmark characteristic of the Tn7-family of transposons. Insertion...Sternberg group’s initial report of VchINT, three separate plasmids and multiple promoters were used to express...Additionally, by expressing multiple crRNAs in parallel (Fig. 3), multiplexed insertions into multiple...
  19. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    Published
    Feb. 13, 2025, 2:15 p.m.
    ... a template, copying the base edit. Figure 2: Cytidine deamination takes place on the free strand...SpCas9 base editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects associated...formation. To further decrease indel formation 1.5-2 fold, the team fused bacteriophage protein Gam to ...which we've discussed in two follow-up posts: (1) (2). Base editors have a narrow editing window of only...K., Arazoe, T., Yachie, N., Banno, S., Kakimoto, M., Tabata, M., Mochizuki, M., Miyabe, A., Araki, M.,...., Slaymaker, I. M., Yen, J., Young, L. E., & Ciaramella, G. (2020). Directed evolution of adenine base...Schwaegerle, N. D., Peraro, L., Young, L., Lee, S.-J., Ciaramella, G., & Gaudelli, N. M. (2023). Improved cytosine...
  20. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    Published
    June 24, 2020, 5:45 p.m.
    ...form or as ready-to-use lentivirus. Figure 2: dCas9-Tet1 demethylates cytosines at promoters and...pX330a dCas9-LSD1 for targeted removal of H3K4me1/2 and H3K9me2 histone modifications. Like the TALE-LSD1...approach for targeted DNA demethylation. Cell Discovery 2: . https://doi.org/10.1038/celldisc.2016.9 Find plasmids...could identify the target genes of previously uncharacterized enhancers. The popular TALEN-based LITE system...
  21. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    Published
    May 7, 2020, 1:15 p.m.
    ...NGG-3′ (“N” can be any nucleotide base, followed by 2 guanines). This limits the use of ABEs as DNA sites... in the promoter region of gamma hemoglobin (HGB1/2) leads to continuous expression of fetal hemoglobin...found that the smaller version, ABE8.x-m, showed comparable editing behavior to ABE8.x-d. Overall, ABE8s ...MS, Rinaldi C, Slaymaker IM, Yen J, Young LE, Ciaramella G (2020) Directed evolution of adenine base editors...
  22. 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...possible to make a timer system composed of two separate FPs (one that matures quickly and the other slowly...s directed mutagenesis also produced the three separate timers shown in the table: Fast-FT, Medium-FT,...varied blue-red maturation times (Table 1). These separate proteins should be useful for monitoring cellular...
  23. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    Published
    Sept. 24, 2024, 1:15 p.m.
    ...of ~75% in response to the magnetic field (Figure 2); they named this variant MagLOV. Andrew was thrilled...’t need any added cofactors.      Figure 2: Fluorescence intensity and ΔF/F of MagLOV, a variant... that don’t even exist yet? Andrew York, Maria Ingaramo, and their team at Calico Life Sciences recently...across. Video courtesy of Andrew York and Maria Ingaramo. MagLOV shows a wealth of potential, and while...Lazzari-Dean, J. R., Lefebvre, A. E. Y. T., York, A. G., Ingaramo, M. (2024). Magnetic control of the brightness...
  24. 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...movement in an open field box or a circular corridor apparatus using a camera or tracking software. Locomotion...the open (risky). Taking advantage of these characteristic traits, rodents can be used to study anxiety...measuring the time spent in a particular part of the apparatus, it turned out that the IL-1 beta overexpressing...
  25. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    Published
    Feb. 11, 2021, 2:15 p.m.
    ...plasmid kit consists of 20 plasmids encoding 14 alpha, 2 beta, and 4 gamma subunits of the heterotrimeric G...activation via bioluminescence resonance energy transfer 2 (BRET2). Check out our blog post on GPCR signaling...created an open source resource called PRESTO-TANGO (parallel receptorome expression and screening via transcriptional...
  26. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    Published
    Dec. 8, 2022, 2:15 p.m.
    ...11 and Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity. Cas7-11 does not display collateral...Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016...science.aav4294 Ozcan, Ahsen, Krajeski, Rohan, Ioannidi, Eleonara, et al., Programmable RNA targeting with the single-protein...
  27. 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 ...buffer. You’ll want to run long enough to get good separation in your ladder, particularly in the sizes around...
  28. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
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    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...not only believe in the separation of church and state, but also the separation of fact and fiction. Our...
  29. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    Published
    Aug. 15, 2017, 1:24 p.m.
    ... fluorescent proteins expressed in the specimen.  2. Fluorescence Activated Cell Sorter (FACs) A machine...photostability of your fluorescent protein when using 2-photon microscopy as this hasn't been well reported...PMID: 18703742. PubMed Central PMCID: PMC2606057. 2. Chtanova, Tatyana, et al. "Dynamics of neutrophil... tagged with a fluorescent protein or dye. It separates the cells mechanically in a vibrating nozzle, ...Schaeffer, Marie, et al. "Dynamic imaging of T cell-parasite interactions in the brains of mice chronically...
  30. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    Published
    July 15, 2025, 1:15 p.m.
    ...short-read sequencing are highlighted in Figure 2. Figure 2: Overview of short-read sequencing technology...synthesized (Figure 1). Sanger sequencing requires four separate reactions, where a fraction of one of the four...The resulting DNA fragments are pooled and then separated by size using electrophoresis, typically capillary...deconvoluted to identify the bases (and, if necessary, separate different samples), the resulting data, or reads...nucleotide that passed through based on these characteristic drops. Figure 3: Overview of Nanopore...To address this issue, the QC team developed a separate script for manually inputting known innocuous ...
  31. 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...controls for an IP. When possible, perform the IP in parallel with both a sample known to express the protein...identify any non-specific interactions, perform parallel IPs with beads only and an isotype control antibody...
  32. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    Published
    Nov. 10, 2016, 3:30 p.m.
    ...diffusion test or Kirby-Bauer susceptibility test (Figure 2), and requires at least 18-24 hours to obtain results...." London: Wellcome Trust & HM Government (2016). 2. CDC Information Pages on Antibiotics 3. WHO Antimicrobial...antibiotic’ can also be used to describe anti-parasitic drugs and anti-fungal drugs, because ‘antibiotic...Resistance can be intrinsic, meaning a basic characteristic that naturally occurs in the bacterium, and...
  33. Deaminet 2026: Breakthroughs in Base Editing, Deaminase Biology, and Therapeutic Translation

    Type
    Blog Post
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    March 10, 2026, 1:14 p.m.
    ...pink. Reproduced from Arantes et al. (2024) under a CC BY-NC 4.0 License.  2. Precision and safety... a new generation of deaminases. Figure 2: Structure of ABE8e*A showing dimerized TadA subunits...191–202. https://doi.org/10.1038/s41586-025-09732-2 Richter, M. F., Zhao, K. T., Eton, E., Lapinaite, ...Lab, Harvard) is a 96-well platform for parallel characterization of hundreds of base editor proteins across...enzyme engineering is accelerating. In parallel, newly characterized editing outcomes are driving the need... critical for its high deamination efficiency (Arantes et al., 2024). By comparing conformational selection... modality‑matched approaches to off‑target characterization. beCasKAS: Linking Cas9 binding to editing...
  34. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...due to the presence of additional helper sequences (2-3). As alluded to above, termination and polyadenylation...and translation processes as well.   Figure 2: The conserved eukaryotic polyadenylation signal directs...transcriptional read-through. Chris Voigt’s lab has characterized a set of prokaryotic terminators and deposited...
  35. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    Published
    Aug. 3, 2023, 1:15 p.m.
    ...from another within a fully stained sample. Figure 2 is an example of the emission spectra for two fluorophores...another during spectral unmixing.     Figure 2: The full emission spectra for BV421 and BV711 on ...experiment, it is important to consider how many parameters you want to use in a given experiment. For very...
  36. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    Published
    Aug. 5, 2025, 1:15 p.m.
    ... follow the same general cyclical process (Figure 2). The differences typically come in when generating...starting point for the next cycle. Figure 2: General process of antibody directed evolution. Created...module_attribute "schema_version" is_json="true" %}{% raw %}2{% endraw %}{% end_module_attribute %}{% module_attribute...antibodies developed by IPI are extensively characterized and validated, with the application data from...
  37. 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...cooperatively to separate nearby AT-rich regions of DNA. In a test tube, the DNA helix can be separated by DnaA...ATP-bound DnaA, the proteins cooperatively can separate the DNA strands. Created with BioRender.com. ...
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