We narrowed to 794 results for: Mpl

... the antibodies time-sensitive, using precious samples, or being compared to other data in a manner that...conjugate that can be activated to emit light with a simple kit, and is readily available on any number of ...ain’t broke, don’t fix it. HRP is easy to find, simple to use, not sensitive to light in its inactive ...detected in the same assay. This is one of the most complex subjects in antibody-based protocols - but thankfully... this is an Antibody 101 post, so we'll keep it simple. If you follow the two basic guidelines below, ...compensation.    Working with antibodies can get a bit complicated, but hopefully this helps you successfully select...

...In complex metazoans, rapid cell division and large scale cell mobility are essential processes during... required for a growing organism to make the complicated transition from a clump of cells to a fully differentiated...organism. Studying cell mobility within these complex environments has never been easy and, for many ...in thick specimens. See the video below for an example of two-photon intravital microscopy from PLoS Pathogens...scientists could easily track GFP expressing cells. For example, by harvesting lymph nodes at different times after...dynamics in lymphoid organs. In lymph nodes, for example, two-photon microscopy allowed scientists to better...time. Photoactivable fluorescent proteins, for example, have been successfully used to study B and T cell...

... of fluorescent dyes/ligands Figure 1: Example structures of fluorophores, fluorescent dyes, and...attached to self-labeling proteins. Some common examples of these self-labeling proteins are SNAP-tag® ...of protein substrates/peptides and antibodies. Example applications of this include coupling to an antibody...photons absorbed over the photons emitted. For example, DAPI is much dimmer than Alexa Fluor 488, and ...cells without causing undue toxicity. A prominent example is DAPI, a dye that binds specifically to the AT-rich...fluorescence-based techniques in biotechnology. Recent examples of advancements include the Janelia Fluor(R) (...finding the right dye for your experiment more complicated. We hope this post helps you in your search for...

... on excitation power in a linear fashion. This implies that reducing the power 2-fold does not reduce ...fluorescent proteins can be directly compared. An example of a photostability measurement of various cyan...results in a loss of fluorescence, which, in its simplest form, can be described by a mono-exponential decay...intensity decay of a fluorophore does not follow a simple mono-exponential decay (Shaner et al, 2008; Bindels...applications in which far less power is used. For example you might find that you can get useful data from...fluorescent protein. In figure 1 we provide an example of such a comparison between cyan fluorescent proteins...

... The destination vector contains sites with complementary overhangs that direct assembly of the final ...plasmid (Option 1) or added using PCR-based amplification (Option 2). Advantages of Golden Gate cloning... reaction. For detailed Golden Gate protocols, complete with helpful tips and tricks, see The Sainsbury...is to "domesticate" your fragment: PCR-based amplification can be used to create silent point mutations...system is an incredibly powerful tool for cloning complicated constructs in a single, high-efficiency step....step. References: Sequential amplification of cloned DNA as tandem multimers using class-IIS restriction...

... features of the gene intact. Here, we show an example gene targeted for tagging with GFP at the C terminus...interest. For a C-terminal tag, as shown in the example above, we inserted a linker and GFP tag at the ...percent of cases, the stem cell will seek out a template for the repair process. By flooding the cell with...using the introduced donor plasmid as a repair template. This results in the insertion of the GFP tag ...tag precisely where we need it, in this example, at the C-terminus of the gene of interest. To get the molecular...CRISPR/Cas9 (Cas9 protein and the crRNA-tracrRNA complex) and the donor plasmid into the cells we use electroporation...

...plasmid are not always the best for others. For example, in our hands we find that a double harvest approach...larger genes such as Cas9. See figure 1 for an example of how changing transfection ratios can affect ... For some transgenes, transfection optimization simply is not enough to get a sufficient titer; this is... volume than with the unconcentrated prep. For example, at Addgene we typically concentrate 100-fold meaning...volumes of lentivirus routinely, you may opt for simply increasing the number of 10 cm2 dishes. While this...available, cost and downstream applications. For example, while the PEG precipitation method is relatively...

... or Cas nickase)  a guide RNA that targets the complex of TadA and dCas to the genomic DNA of interest...effect of removing the wild-type TadA from the complex the researchers developed a second set: The ABE8... the team replaced S. pyogenes Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes... ability to use non-standard Cas9 in the ABE8 complexes enables a broader targeting scope for base editing... the corrected cells back to the patients. For example, defects in hemoglobin (𝛽-globin), which cause...that require high DNA editing specificity, for example for potential therapeutic approaches, the authors...

...the ribosome complex and Dr. Steitz quipped that RNA in the ribosome is in a complicated conformation ...Lowe lab have allowed them to begin understanding complex regulation of and by tRNAs and tRNA-derived RNAs...long hypothesized that life started with RNA and a simple cell membrane. His recent work shows that they ...replication with no proteins). The chemistry is complex and this work requires an interdisciplinary team... they do it. Of course, No conference would be complete without CRISPR Michael Terns from of the University...

...are often thought of as simple organisms , they have many levels of complexity, especially when trying...learning how to grow and care for the worms. As an example of one of the problems we work collaboratively ...We attempt to deconstruct and understand this complexity using existing C. elegans mutants. For instance...data is freely available, and the collection of samples and sequencing is being organized and funded by...Kittybiome project gives them access to rich and complex microbiome data for analysis. And, well, cats.....

... millions of people die from cancer-related complications each year. Despite these grim statistics, new...cancer, really? While cancer is extraordinarily complex at the molecular level, at the cellular level there...cancer research has focused on picking apart the complex cellular pathways involved in the development of...pathway. Its major appeal is that it enables the simple construction of vectors to study the function of...transcript most commonly found across all TCGA tumor samples is represented, making the results of any such ...biggest challenges facing researchers today is the complexity of the pathways involved in the development of...

...realized how perfectly flat the complex is on this surface”….where the complex is docked on the membrane of...like “Oh,…I didn’t quite appreciate that….” For example, when Tom Steitz was presented with a physical ... close to its reported cleavage site on the complementary DNA strand. And this is why we love working ...cleavage”. Jacob computationally added the non-template strand to the Jinek structure so that the resulting...double-stranded DNA, recognize a PAM sequence, allow the complementary base pairing between the guide RNA and the target...

...though three examples depend on many specific details about the individuals, each example provides a scenario...machine? Let’s walk through a more specific example. Perhaps you and your lab mate are negotiating ... something valuable. Importantly, as the third example shows, the ultimate resolution can involve ideas...your organization. In my non-academic roles, some examples of negotiation have been: A job offer conflicts...explore while also ensuring that my work gets completed? I want my company to pay for me to attend a conference...

...endogenously expressed SCF complex members, an E3 ubiquitin ligase complex. Finally, when auxin is added...fusion protein with the endogenous CRBN E3 ligase complex which will polyubiquitinate the fusion protein....target protein-of-interest and the CRBN E3 ligase complex leading to ubiquitination and proteasomal degradation...acid (IAA), the TIR1 F-box associated E3 ligase complex will ubiquitinate the AID-tagged protein which ...Inducible, reversible system for the rapid and complete degradation of proteins in mammalian cells. Proc...

...allowing very low background level of fluorescence. Examples for such photoactivatable or photoconvertible ... and their emission is green or red light. For example, T-Sapphire, LSSmOrange, and LSSmKate. Fluorescent...sensor), and iGluSnFr (glutamate sensor). More examples of these biosensors can be found at Addgene. ...also is also called Bimolecular Fluorescence Complementation (BiFC). 8 points to keep in mind when choosing...system can excite, and detect the emission. For example, if your microscope has only two lasers, at 488nm...minutes after it is translated to a few hours. For example, superfolder GFP (sfGFP) and mNeonGFP can fold ...

... takes 1-1.5 hours to complete, while this syringe-based method can be completed in 20 to 30 minutes. ... centrifugation. At this point, the plasmid DNA sample can be treated with RNase to remove RNA contamination...directly into a PCR buffer, where DNA or RNA can be amplified. Compared to a commercial magnetic bead-based ...waste footprint. The Zhao and Deng labs developed a simple method for regenerating silica columns that come... reduce the number of spin columns you need to complete your research. Do you know of other nucleic acid...

...major findings and then focusing more on broader implications rather than diving deep into the individual ...carefully chosen for each group and topic. For example, it may be useful to adapt the second version (...stay abreast of current trends in the literature, sampling a breadth of work from a field (perhaps even two...to allow those who could not attend a meeting to simply stay caught up and to allow those who may be interested...get feedback from those who didn’t attend. For example, we got a good bit of early feedback from those...own knowledge of the field. Perhaps interest has simply waned. When attendance slumps below a certain level...

...mediate cleavage rather than the multi-subunit complex employed by class 1 systems. A putative new class 2...nucleases. Using Francisella Cpf1 (FnCpf1), they employed an E. coli plasmid depletion assay to discover...than following it. The Cpf1 crRNA is also much simpler in structure than Cas9’s; only a short stem-loop...CRISPR systems based on Cas9 were thought to be the simplest CRISPR systems and the easiest to adapt to genome...confirmed. To prevent new editing post-HDR, repair templates should remove the PAM sequence. Comparing Cpf1...

...Its RNA-guided nature makes CRISPRa much simpler to implement in genome-wide screens over cDNA libraries... Rochester Medical Center. During development, complex genetic programs specify and assemble diverse arrays...genetic underpinnings of these processes has been complicated by the lack of precise genetic tools for modulating... must be screened to identify effective ones. Simplified in vivo delivery using SPH mice CRISPRa has largely...population, as expected. Although this strategy was employed to study neuronal differentiation, it could also...

... The central nervous system (CNS) orchestrates complex processes enabling organisms to control their movements...photostable than GFP. In addition, antibody-based amplification of their signals is weak and increases background...sites for primary antibodies which can be used to amplify the direct smFP signal within sub-cellular structures...used to distinguish highly similar proteins. For example, N-cadherin (cadherin-2) is a postsynaptic cell...super-resolution STORM imaging, and electron microscopy. Sample preparation for these techniques sometimes makes...smFPs retain their ability to be detected after sample preparation better than standard proteins tags ...