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We narrowed to 594 results for: cel.2

Showing: 341 - 360 of 594 results

  1. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    ...26000849. 2. Grimm, Jonathan B., et al. "A general method to improve fluorophores for live-cell and single-molecule...expressed as a fusion with your favorite protein and (2) a synthetic fluorophore-contianing “ligand” that ...). These cell-permeable Janelia Fluor (JF) dyes have excellent properties inside living cells, especially... fluorescent proteins. In-cell labeling strategies The majority of in-cell labeling strategies have two...and oligonucleotide labels for fixed-cell imaging, not for live-cell applications. Thus, these fluorophores..."Imaging live-cell dynamics and structure at the single-molecule level." Molecular cell 58.4 (2015): 644...sparked a renaissance in biological imaging. Suddenly, cell biologists were no longer beholden to chemists and...

  2. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...suppress Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi.org/10.1093/mutage...antibiotic selection. Essentially, this gene is a 2-in-1 providing both bacterial and eukaryotic selection...prevent cell death. One of the cool things about amp is that it's exclusively toxic to dividing cells. The... continuous cellular processes (e.g., protein synthesis), while amp only affects the cell wall synthesis...Check icon" %} Spectinomycin Excellent No Just bacteria Not SHuffle cells {% icon icon_set="fontawesome...icon" %} Zeocin Excellent No Bacteria, plants, and eukaryotes Not Tn5-containing cells {% icon icon_set..., penicillin, and is a beta-lactam that inhibits cell wall synthesis. The AmpR (amp resistance) gene produces...

  3. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...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...to monitor cell, organelle, and protein localization. For instance, you might dye your cells and look at...either in whole cells or as a pool of fusion proteins localized to a specific region of the cell. For example...the relationship between the age of a cell, protein, or cellular structure and a particular biological...trafficking to a subcellular location, start of gene expression, development of a cell structure, etc)....timers, researchers can look at a single image of a cell and understand how protein localization changes ...

  4. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ...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....and yeast cells are cost-effective and faster-growing than mammalian cells, but mammalian cells can perform...your cells with your plasmid(s), and then wait 7-14 days (in mammalian cells) to collect the cell culture...production system, cell lines typically used include Human Embryonic Kidney (HEK) cells and Chinese Hamster...recombinant antibodies with some molecular biology and cell culture experience. Let’s go over the basics of ... place in bacterial, yeast, plant, or mammalian cells. Each of these systems has their advantages and ...

  5. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...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...fluorescence activated cell sorting (FACS) to isolate differentiated from undifferentiated cells at the end of ...these cell types are used. The lack of good flow cytometric antibodies for many CNS-resident cell types...that drive neuronal fate and reprogramming." Cell stem cell 23.5 (2018): 758-771. PubMed PMID: 30318302...CRISPRa screening approach in embryonic stem (ES) cells to determine which transcription factors promote...antibodies, downstream of the TUJ1 coding sequence. ES cells that stably express the dCas9-Suntag activator were...

  6. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...retain their individual activities when separated (2), and that DBD and TAs from different systems could... Enzymol. 1983;100:293-308. PubMed PMID: 6312261. 2. Keegan, L., Gill, G., and Ptashne, M. Separation ...entire proteome of a particular cell type, or a subset based on a cellular compartment or growth stage)....extensively to screen for molecular interactions in the cell, including protein-protein, protein-DNA and protein-RNA...field of eukaryotic transcriptional activation and cell biology. During this period, proteins were successfully...The reporter gene then allows detection of those cells where binding between the proteins of interest has...alternative fungi (C. albicans, pC2HB) (7) and mammalian cells. If the problem is that your proteins are not post-translationally...

  7. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...maybe label) your sample (i.e. cultured cells or tissues).  Step 2: attach anchors to your sample that will...quite so much. Created with biorender.com Steps 2-4 are not found in standard IHC/ICC protocols and ... for EM is not something you find in the average cell and molecular biology lab, and EM is limited in ...ultrastructural relationships that influence everything from cell division to memory, but don’t have access to the...properties of the sample itself. For example, cultured cells are typically easier to expand than mouse brain ..., Mohar B, Eddison M, et al (2022) Visualizing cellular and tissue ultrastructure using Ten-fold Robust... Zwettler FU, Guennec ML, et al (2019) Imaging cellular ultrastructures using expansion microscopy (U-...

  8. An Interview with Connie Cepko - Gene Therapy, Plasmid Tools and Insights to Success

    Type
    Blog Post
    ...from this publication at Addgene. 2. Tang, Jonathan C.Y., et al. “Cell type-specific manipulation with ...fluorescent proteins as scaffolds for cell-specific gene manipulation.” Cell 154(4) (2013): 928-939. PubMed ...Cepko Interested in Working on the Eye 9:05-10:16 : Cell Targeting in Retinal Gene Therapies 10:17-14:34 ...28:39 : Methods for Delivering Multiple Genes to Cells for Gene Therapy 28:40-35:10 : GFP Activatable Tools...

  9. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    ...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... attack the cell wall of bacterial cells; however, Mycoplasma species do not possess cell walls. As a ...bacteriophage (viruses that only infect bacterial cells), or a bacterium picking up resistance genes from...conserved, non-negotiable process within the bacterial cell, antibiotic resistance is very likely to develop...

  10. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ... 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...that it appears “newly synthesized” to the cell. Thus, the cell repairs the DNA using the U-containing strand..., which may lead to cell death rather than NHEJ repair, thus removing these cells from the edited population...target modification in primary T cells, making them a promising tool for cell therapy applications. Dual base...

  11. Antibodies 101: Choosing the Right Isotype

    Type
    Blog Post
    ... reduce the background staining (Table 2).      Table 2: Secondary antibody selections for a four-antibody...for Light and Electron Microscopy. In Methods in Cell Biology (pp. 283–310). Elsevier. https://doi.org...

  12. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    ...enables efficient knockout (KO) of genes in immune cells; (2) is not immunogenic, a necessity for in vivo studies...activated T cells, precluding analyses of T cell differentiation beginning with naïve T cells. The Framework...screens have been used to interrogate T cells, an important immune cell type, in more complex and physiological...naïve T cells. More broadly, this system can be used to perform in vivo screens in other cell types. A...screens to assess T cell responses and clonal dynamics in lymph nodes (LN) and tumor cells. Figure used with...FITS In vitro CRISPR screens in immune cells have identified many putative regulators of immunity. However...screens cannot faithfully model the complex spatial, cellular, metabolic, and cytokine microenvironment that...

  13. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ...ability to easily cure cells of the plasmid after integration. Figure 2: (A) Streamlined VchINT...et al., 2020). Genomic targets require an upstream 2-bp PAM sequence for recognition by QCascade; although...into the mini-transposon Transform plasmid(s) into cells and plate overnight Screen colonies phenotypically...distinct biological elements, CRISPR-transposons excel at programmable insertions of even very large genetic...

  14. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ...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... . Steps of an IP: 1) Protein extracts from cells or tissues contain a complex mix of proteins seen...common immobilization methods utilizes bacterial cell wall proteins, Protein A and Protein G, which bind...does not. Common negative controls include knockout cell lines or tissues that do not express the protein...

  15. 5 Science Rap Videos & Their Creator: Tom McFadden

    Type
    Blog Post
    ...important role in solving the structure of DNA. 2. The Immortal Rap of Henrietta Lacks Students in Oakland...the African American woman whose cervical cancer cells became a mainstain of biological study without her...

  17. Plasmids 101: How to Name Your Plasmid in 3 Easy Steps

    Type
    Blog Post
    ...available empty backbones, visit Vector Database. Step 2: Insert name  Include information about the insert...selection markers (for use in other cell types, e.g. eukaryotic cells). pBACKBONE-XXXXX Tip: For a catalog...

  18. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...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...number of cells have been edited. For many experiments, the end goal is to generate a clonal cell line, so...designed your gRNA and introduced it into your target cells with Cas9. Hooray! Now it’s time to make sure your...blog post we’ll explain how to verify that your cells were appropriately edited for your desired mutation...important to know a few things about your target cells and the desired edits. First, you will need to know.... Validating knockout mutations When a knockout cell line is desired, any mutation that introduces a ...

  19. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...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...replicating too quickly can slow cell growth or even kill the host cell. For stringently controlled plasmids...All plasmids rely on their host cell's replication machinery in order to replicate—but not always to ...be synchronized with the replication of the host cell's chromosomal DNA.  Plasmids are said to be under... stringent control when they depend on the host cell’s initiation proteins to start their own replication... occurs at the appropriate time—for example, the cell must have enough nutrients available to complete...

  20. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...sequence is unique compared to the rest of the genome.  2) The target is present immediately upstream of a Protospacer...utilized allowing for PAM flexibility. Fig. 2: Prime editor engaging target DNA. The reverse transcriptase...Generating a no-fuss genetic knockout  Knock-out cells or animals are engineered by introducing Cas9-mediated...transcription by blocking its initiation. In mammalian cells, robust repression requires targeting dCas9 along...study essential genes whose knockout would cause cell lethality. The Krüppel associated box (KRAB) is ...repressing target genes in a variety of species and cell types can be found on Addgene’s website. Activating...activating target genes in a variety of species and cell types can be found on Addgene’s website. ...
Showing: 341 - 360 of 594 results