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We narrowed to 490 results for: ARA-2

Showing: 261 - 280 of 490 results

  1. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ... 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...

  2. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...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. ...

  3. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    ...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 ...

  5. Advice for Starting a Biotech Company from a Grad Student turned Entrepreneur

    Type
    Blog Post
    ...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...

  6. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    ...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...

  7. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...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...

  8. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...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 ...

  9. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    ...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...

  10. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ...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...

  11. 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...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...

  12. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...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...

  13. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    ...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...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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...

  20. 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...
Showing: 261 - 280 of 490 results