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  1. 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...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...
  2. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    ...of ~75% in response to the magnetic field (Figure 2); they named this variant MagLOV. Andrew was thrilled...didn’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...
  3. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ...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...
  4. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    ...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...
  5. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    ...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...
  6. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ... 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...
  7. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    ...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...
  8. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ... 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...
  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...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...
  10. 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...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...
  11. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    ...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...
  12. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    ...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...
  13. 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...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. ...
  14. Zhang Lab's CRISPR Frequently Asked Questions

    Type
    Collection
    ...extraction solution, runs the protocol, then uses around 2-4ul in each 50ul PCR reaction. The gDNA extracted ...system, developed by the Zhang lab, which has comparable efficiency to the optimized chimeric design but...better for PCR and surveyor. I used DNA polymerase Takara Ex Taq ™ DNA Polymerase for my genomic PCR, but...successfully used Herc II or Kapa Hifi in PCR. Maybe this Takara enzyme is not very robust in this case for EMX1...processing of the crRNA) is also expressed from a separate promoter. In the pre-processing form, the crRNA...
  15. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ... 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...10634178. 8. Murnane, John P., Michael J. Yezzi, and Barbara R. Young. "Recombination events during integration...
  16. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    ...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 ...once), however, SAM exhibits activation levels comparable to other popular activation methods (VPR and ...
  17. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ...mean you have one copy of the gene; an RQ value of 2 means you have two copies, and so on and so forth....quantitative PCR. Expert Rev Mol Diagn. 2005 Mar;5(2):209-19. doi: 10.1586/14737159.5.2.209. PMID: 15833050...falciparum multidrug resistance-1 gene (pfmdr1). J Parasitol. 2011 Oct;97(5):939-42. doi: 10.1645/GE-2792.1...
Showing: 281 - 300 of 486 results