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We narrowed to 350 results for: SAM activators

Showing: 301 - 320 of 350 results
  1. Protocol - How to Design Primers

    Type
    Protocol
    ...must selectively block and unblock repeatedly the reactive groups on a nucleotide when adding a nucleotide...primer is used to amplify a eukaryotic genomic DNA sample. However, a primer should not be too long (> 30...
  2. Chemogenetics Guide

    Type
    Guide
    ...For example, PSAM Q79G,Q139G is activated by PSEM 22S , while PSAM L141F,Y115F is activated by PSEM 89S...of Na + and/or K + ions, and activates neuronal activity. Figure 3: PSAM4-based ligand-gated ion channels...ligands, could be activated by synthetic ligands. Specifically, Receptors Activated Solely by Synthetic...constitutive activity, making them less than ideal. DREADDs Designer Receptors Exclusively Activated by Designer...endogenous ligands, have low constitutive activity, and their activating ligands have few, if any off-target...ions, and inhibits neuronal activity. PSAM-5HT3 PSAM-5HT3 LGICs pair a PSAM domain with a 5-HT 3 serotonin...off-target activity, and do not seem to have the same back-metabolism issues as CNO and are attractive alternatives...
  3. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    ...compare different clones of the same target to see if the results are the same/similar, emphasizing the antibody...your sample to. For example, we use our negative control to compare exposure times of our samples. A prominent...confirming it will produce the same results even to a different sample. Finally, as a lab we discuss the...underlying causes of this unreliability: cross-reactivity (binds to non-target proteins), variability (...Baker, 2015), causing a huge waste of precious samples, time, and funds – up to US$350 million in 2015...noise. We then compare the negative control to a sample with the primary included to confirm that the signal...antibody and not from the background noise. If the sample images similarly to the negative control, that ...
  4. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    ...particles = number of particles per mL of sample. The purity of the sample can be estimated by looking at the...Adenoviral vectors (AdV) are attractive vectors for research applications and gene therapy: they can ...Trapnell. "Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy." Journal...
  5. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ...thiols, to cleave disulfide bonds. Load the samples, one sample per lane, onto the top of a resolving gel...all of the proteins in a sample migrate through the stacking gel at the same pace and enter the resolving...consistent size and density present in all samples. Uneven sample loading or incomplete protein transfer ...Figure 1: Overview of the Western blot process. The sample, a mixture of protein is separated in a gel, transferred...is the first step of a western. To prepare the samples for SDS-PAGE, measure the protein content and normalize...normalize to ensure equivalent loading. Denature the samples to their primary amino acid sequence by boiling...crosslinked polymer acrylamide. In addition to the samples, a protein standard of known molecular weight in...
  6. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...known titer on every qPCR run. Note that the rAAV sample has to be digested with DNase before undergoing...Grimm, D., Kern, A., Pawlita, M., Ferrari, F., Samulski, R., & Kleinschmidt, J. (1999). Titration of AAV...Harvey, B. K. (2017). Assaying the Stability and Inactivation of AAV Serotype 1 Vectors. Human Gene Therapy..., J., Hennigs, J. K., & Körbelin, J. (2021). Inactivation of Adeno-Associated Viral Vectors by Oxidant-Based...
  7. Antibodies 101: Validation

    Type
    Blog Post
    ...antibody signal in the retinal sample, but little to no signal in the liver sample, which you do (C), so your...epitopes on the same target reduces the likelihood that both antibodies would bind to the same off-target ...core goals of antibody validation:   Confirm reactivity with the target antigen - Recognition of an immunogen...to - particularly when you start using complex samples such as whole cell lysate or tissue sections. It...require variable expression of your target across samples - it is hard to distinguish between specific, invariable...there is a wealth of transcriptomic data from this sample type, so you use your bioinformatics skills (or...Removing (or reducing) the protein of interest in your sample, should lead to reduced or eliminated antibody ...
  8. Plan Your Experiment

    Type
    Guide
    ...fusion with transcriptional activator (such as dCas9-VP64) Many different activator systems exist, including...including the multi-plasmid SAM system Activate Design and Clone Your gRNA Achieving your desired edit comes...Knockouts Activation/interference Needs good primer design Western blot Knockouts Activation/interference...the gRNA activity. Be sure to sequence the genomic region you intend to target in your samples to ensure...proteins) Many base editors fused to Cas9 exert their activity within a very narrow window proximal to the PAM...dCas9 alone for mammalian cell lines Interfere Activate (CRISPRa) Increase gene expression without permanently...essential protein domain could abolish protein activity and essentially function as a “knockout,” but ...
  9. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ...pseudotype To pseudotype your viral vector, you use the same protocol for generating lentivirus, but you use ...Different pseudotyped viruses are differentially inactivated by nonspecific complement mechanisms in serum...anti-alphaGal antibodies. The VSV-G envelope protein is inactivated by serum regardless of the producer cell type...high concentrations as described above and is inactivated by the complement system in serum (DePolo et ...Vector Particles Produced in Human Cells Are Inactivated by Human Serum. Molecular Therapy 2:218–222 ....
  10. Cultivating Community Science at BosLab

    Type
    Blog Post
    ... DIY Biology and, on occasion, would meet in the same space where BosLab exists today. Back then, it consisted...saw the potential in the space and decided to reactivate it. We started out by holding a few journal club... getting the community to collect various soil samples, extract DNA, sequence it, and analyze the data... characterize the microbiomes of the different samples. The goal is to not only learn something about ...
  11. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...strategies to investigate a gene's function is to inactivate, or "knockout", the gene by replacing it or disrupting...describe the techniques and the plasmids used to inactivate specific genes in mammalian cells. Despite the...one of its exons has simply been replaced with the same sequence flanked by LoxP sites while the resistance... are easier to use and are more efficient at inactivating genes. CRISPR can target a genomic sequence ...
  12. Inside Addgene's 10 Year Anniversary Gala

    Type
    Blog Post
    ... the interactive photo graffiti wall. I had a lot of fun playing the prototype of an interactive game ...playing all night was hand-picked (like our plasmid samples!) by Addgenies. If you were there, you might have...
  13. In Living Color: The Skinny on In Vivo Imaging Tools

    Type
    Blog Post
    ...last year, Verkhusha added two far-red light photoactivatable (PA) near-infrared fluorescent proteins (FPs...signal-to-background ratio, thus, allowing higher resolution in samples containing substantial autofluorescence background...751-4. doi:10.1038/nmeth.2521) Far-red light photoactivatable near-infrared fluorescent proteins engineered...
  14. Plasmids 101: Antibiotic Resistance Genes

    Type
    Blog Post
    ... are trying to disrupt or delete, which both inactivates the gene and acts as a marker for the mutation...of experiments it is best practice not to use the same resistance cassette for the mutation and for plasmid...cassettes and your gene of interest is cloned into/inactivates or completely removes (in the case of Gateway...
  15. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ...approach – co-CRISPR from Craig Mello’s lab – used inactivation of the unc-22 gene as their selection marker...Drosophila S2 cells (Böttcher et al., 2014), inactivation of non-homologous end-joining results in a further...changes 5’ to the PAM, sequences corresponding to the same strand as the PAM produced a higher editing efficiency...elegansGenome with Lethal Mutation Co-Conversion and Inactivation of NHEJ Repair. Genetics 199:363–377 . https...
  16. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    ... have a higher transduction efficiency, i.e. the same number of AAV particles is able to infect more cells... protects them from antibody recognition and inactivation (György et al., 2014), but scientists have yet...difficult to reach, and are more resistant to inactivation through anti-AAV antibodies. References György...
  17. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    ...classic approach for studying protein turnover is radioactive pulse-chase experiments. However, these experiments...require radiolabeled amino acids. The use of radioactive materials is highly regulated, requires designated... suspected to interact, are added to a cellular sample. Both antibodies have a short antibody specific...
  18. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    ...This can differ between fluorescent proteins in the same spectral class (Cranfill et al, 2016). This non-...described by a mono-exponential decay (analogous to radioactive decay). Consequently, the intensity decay is ...comparison of several fluorescent proteins of the same spectral class on the microscopy system and in the...
  19. A Guide to Designing a Scientific Poster: Content, Layout, and More

    Type
    Blog Post
    ...your poster. While a poster generally contains the same sections as a primary research article, it’s important...can be accomplished with an exciting title and attractive figures. Next, the poster should be legible from...four column style, and in all cases the idea is the same; the reader will navigate the poster from column...
  20. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    ...under the control of an inducer, i.e. tet-off transactivator (tTA), is injected at a high dose to maximize...An AAV expressing the inducer (i.e. tet-off transactivator) is co-injected. The dose of this AAV can be... Central PMCID: 5088052. 3. Lentz TB, Gray SJ, Samulski RJ. Viral Vectors for Gene Delivery to the Central...
Showing: 301 - 320 of 350 results