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Showing: 271 - 300 of 707 results
  1. Antibodies 101: Avoiding the Mouse on Mouse Mess in IHC

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    Blog Post
    ...immunocytochemistry (ICC). Not all of the solutions described here may be as effective for other techniques,...Technique Adaptable to Biotin-Free Reagents, Immunofluorescence, and Multiple Antibody Staining. J Histochem...Addgene blog Antibodies 101: Introduction to Immunofluorescence Antibodies 101: Affinity Reagents Antibodies...
  2. Three Tips for Preventing Viral Plasmid Recombination in Your Samples

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    Blog Post
    ...instability-prone lentiviral vector plasmid in Escherichia coli Stbl3. 3 Biotech 3:61–70. https://doi.org...lentiviral expression vectors using reduced-genomeEscherichia coli. BioTechniques 43:466–470. https://doi.org...cloning strategy for efficient genome editing in Escherichia coli. Microb Cell Fact 19. https://doi.org/10.1186...
  3. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

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    ...our AAV preparations. We recently published a description of the VGS process and use cases in Human Gene...ligation and tagmentation preparation approaches described previously require a preliminary double stranding...signature sequences in the data. Unlike previously described thermodynamics-based methods of serotype confirmation...
  4. The Importance of High Titer for AAV Transductions

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    Blog Post
    ... light responsive than fluorescent proteins required to get a good fluorescent signal. It may not be necessary...are often necessary to visualize expression of fluorescent proteins, for example, as many viral particles...
  5. R Bodies: Membrane-Rupturing Microscopic Tools

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    Blog Post
    ...bodies decorated with a fluorescent protein remain functional. Attaching the fluorescent protein fusions allowed... body dynamics. Second, we showed that these fluorescent R bodies can rupture E. coli spheroplasts (cells...
  6. Synthetic Photobiology: Optogenetics for E. coli

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    Blog Post
    ...Refactoring and optimization of light-switchable Escherichia coli two-component systems. Schmidl SR, Sheth... at Addgene. Synthetic biology: engineering Escherichia coli to see light. Levskaya A, Chevalier AA, ...Multichromatic control of gene expression in Escherichia coli. Tabor JJ, Levskaya A, Voigt CA. J Mol Biol...
  7. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

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    Blog Post
    ...Harvard University. In my previous two posts, I’ve described the fundamentals of how to work with Drosophila...Drosophila as an experimental model organism. I then described the Gal4/UAS system used by geneticists to study...manipulations. You can produce some very beautiful immunofluorescent images like in this paper (2). Metabolic assays...
  8. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

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    Blog Post
    ...positive effect on cancer treatment (Ref 1). As described in the presentation, starvation or the use of ...can find useful tips on in vivo imaging in our Fluorescent Protein Guide. Throughout the conference aspirin... with T cell receptor engineering (Ref 5)  as described by Blue Flame Addgene depositor Professor Michel...
  9. Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection

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    Blog Post
    ...27478040. The lung cancer mutant collection is described in this paper. Systematic Functional Interrogation...the genetic pathway reference collections are described in this paper Systematic morphological profiling...  The genetic pathway reference collection is described in this publication We also wrote an extensive...
  10. Antibodies 101: Flow Cytometry Controls

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    Blog Post
    ...cytometry controls we’ll discuss are single color, fluorescence minus one (FMO), and isotype. You should always...controls like FMO or isotype controls, which we’ll describe a bit later. Single Colors Single color controls...(C) with APC-Cy7.   FMOs FMO stands for fluorescence minus one. It is used as a negative control ...
  11. Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification

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    Blog Post
    ...protein. OptoNBs were also tagged with an infrared fluorescent protein tag which let the team detect its co-...revealed a blue light-dependent shift in mCherry fluorescent to the surface of the bead, suggesting that OptoNBs...OptoNB-SOS activated Ras/Erk signaling by using a fluorescent reporter for Erk kinase translocation. These ...
  12. GCE4All: Making Genetic Code Expansion Accessible

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    Blog Post
    ...biologically relevant modification, or add in fluorescent or chemically reactive side chains, all of which...through a seminal paper from Peter Schultz’s lab describing a technique that allowed for intentional incorporation...of Phosphoserine into Recombinant Proteins in Escherichia coli. 12(21). https://doi.org/10.21769/BioProtoc...
  13. Kiran Musunuru on the Newest TALEN Genome-Editing System

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    Blog Post
    ...Cell, Musunuru, Chad Cowan and their colleagues describe a much more efficient tool for doing that disease...people unfamiliar with this new TALEN kit, can you describe it for us? Musunuru: TALENs are a type of genome...including FokI endonuclease domain as well as fluorescent proteins used to sort the cells after the plasmids...
  14. Antibodies 101: Conjugation

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    Blog Post
    ...either through a chromogenic or chemiluminescent reaction. Chemiluminescence, which is light produced by ...detect your antibodies using techniques like fluorescence microscopy, flow cytometry, or spectrophotometry...
  15. 5 Tips for Troubleshooting Viral Transductions

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    Blog Post
    ...should appear fluorescent if they were infected. While there may be some low-level fluorescent protein or...virus. Alternatively, if the virus encodes a fluorescent protein, the packaging cells could be examined...
  16. Plasmids 101: Modular Cloning

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    Blog Post
    ...specifically refers to the organizational schema described above, and not any particular cloning technique...cellular engineering goal, several of which are described below. Multigene circuits assembled through MoClo...relationship between genotype and phenotype is best described with “it’s complicated.” Applying MoClo Interested...
  17. Troubleshooting and Optimizing a Western Blot

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    Blog Post
    ...colorimetric ladder, visible to the naked eye; a fluorescent ladder compatible with your imaging system; or...having to re-run the western.  Remember that chemilumescent methods, like horseradish peroxidase protein...Underexposed Increase exposure time Chemilumenescence: use a more sensitive exposure kit Too...Overexposed blot Shorter exposure time Chemilumenescence: use a less sensitive exposure kit No ...
  18. Hot Plasmids: Winter 2025

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    Blog Post
    ...photostable than fluorescent proteins, making them ideal for single-molecule fluorescence techniques. After...used to measure signal along the dendrite. D) Fluorescence signal as a function of the distance between...
  19. Plasmids 101: Knockout/Knock-In Plasmids

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    Blog Post
    ...template with the endogenous gene. Here, we'll describe the techniques and the plasmids used to inactivate...Conditional knockouts The methods and plasmids described in the preceding sections are simple ways of knocking...knockout, researchers often use the Cre/Lox system described earlier. In this case, you design your targeting...
  20. Cloning Mammalian Cells with the Agarose Method

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    Blog Post
    ..., if your protein is fused with a fluorescent protein), fluorescence microscopy can theoretically be used...Addgene.org Check Out Our Protocols Pages Browse Fluorescent Protein Plasmids Find Mammalian Expression Vectors...
  21. Plasmids 101: 5 factors to help you choose the right cloning method

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    Blog Post
    ...following cloning methodologies have been extensively described in other articles; so if you are interested in... to assemble up to 6 different fragments in a prescribed order. These fragments can be pooled in one tube...homology. You might also consider FastCloning, first described by Chaokun Li and colleagues. Here you simply ...
  22. Plasmids 101: Degron Tags

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    Blog Post
    ...proteasome for degradation. Like several other tags described here, the dTAG uses a small molecule, a tag, and...for visualization of proteins of interest. A fluorescent Halo ligand can be added to media, buffer, etc...there are antibodies to it and ways to achieve fluorescence with it. AID The auxin-inducible degron (AID...
  23. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

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    Blog Post
    ...organism carrying a synthetic gene drive were to escape the laboratory, the drive could eventually spread...protocols, they can reduce the risk of accidental escape to a negligible level. One major reason we chose...performing experiments in a geographic area where escaped organisms won't be able to find mates2. For example...
  24. AAVs CREATed for Gene Delivery to the CNS and PNS

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    Blog Post
    ...are easy to trace and not lost in a jumble of fluorescent neurons. A simple but flawed approach to multi-color...constitutively express a red, green, or blue (RGB) fluorescent protein (4a-c). When high doses of this viral...morphology studies when used as part of a two part fluorescent tag-then-express system that allows for controlled...
  25. Transferable Skills Guide: Identifying Your Transferable Skills

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    Blog Post
    ...looking at your resume while they wrote the job description. You check off all the boxes - you have the desired... gaps in scientists' transferrable skills. In describing the limitations of the study, the authors noted...over email. Examine your current and past job descriptions. Scan through all the things for which you are...
  26. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

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    ...worked extensively with enChIP systems. Can you describe this technology and its advantages? Fujii: In ...enChIP system using CRISPR technology. Can you describe this technology and some of its potential applications... any other technologies that you would like to describe?  Fujii: I believe that "Biology is Method", i.e...
  27. "Hall of Fame" AAV Enhancers from the Allen Institute for Brain Science

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    Blog Post
    ... a minimal promoter to drive expression of a fluorescent gene and packaged into viral vectors that can... To evaluate the resulting expression of the fluorescent reporter, the brain tissue was extracted, sectioned...includes annotated images of the brain sections describing the labeled regions, as well as the strength ...
  28. Plasmids 101: Codon usage bias

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    Blog Post
    ...every amino acid I’d like to encode,” but, as described above, not every region of a protein should necessarily...rate dependence of transfer RNA abundance in Escherichia coli." The EMBO journal 9.13 (1990): 4359-4366...rate of full‐length human proteins expressed in Escherichia coli." Protein Science 19.7 (2010): 1312-1326...
Showing: 271 - 300 of 707 results