...wages, overtime tracking? What are institutions doing? Institutions are now preparing to comply with the...resource on what institutions are actually doing (using the list of 340 institutions with postdocs in ...In 2014, 89% of institutions had a minimum salary policy: of those, 51% of institutions set their postdoctoral...should be on the lookout for what their institutions, or institutions they're thinking about applying to, ...salaries will have to double at some institutions; and that institutions which can more easily provide bridge...This post was contributed by Future of Research Executive Director, Gary McDowell. On Dec 1st, the threshold...scientists and those in technical roles. Therefore, institutions are responsible for ensuring that either all...

...were at institutions focused only on tracking hours, and 22% of postdocs were at institutions where no... of some public institutions) to cancel plans to raise salaries. What are institutions doing after the...planning unionization at their institutions. One clear message that institutions have sent, with the lack of...their institution is doing from Future of Research over social media, not from their institution), is ... contributed by guest blogger, Gary McDowell, executive director of Future of Research. On December 1st...believe that, while the mandate has been removed, institutions should still go ahead with raises, as for years...of academic stakeholders. However, while most institutions are indeed going ahead with their planned raises...

...those in eukaryotic cells. Some promoters are constitutively active and on all the time while others are...expression promoter from the human cytomegalovirus Constitutive  May contain an enhancer region. Can be silenced...expression from human elongation factor 1 alpha Constitutive  Tends to give consistent expression regardless...promoter from the simian vacuolating virus 40 Constitutive  May include an enhancer. PGK1 (human or mouse...promoter from phosphoglycerate kinase gene.  Constitutive  Widespread expression, but may vary by cell...Mammalian promoter from the human ubiquitin C gene Constitutive  As the name implies, this promoter is ubiquitous...mRNA Mammalian promoter from beta actin gene Constitutive  Ubiquitous. Chicken version is commonly used...

...of that antibody dilution. Adjust the antibody dilution until your protein dilutions are in the dynamic... get it to work well. Dilutions Most antibodies come with a recommended dilution and/or range (typically... the antibody at the recommended dilution. If the antibody dilution’s dynamic range encompasses your protein...consider a dilution range from 1:100–1:1,000. If your antibodies come from ascites fluid, try a dilution range...two types of gradients. Protein gradients use dilutions of your sample or protein of interest to understand...reassembled and imaged. In this example, a 1:1,000 dilution gives a high signal with low background. Created...running properly but you’re not seeing sufficient resolution in the bands, you may want to consult Tables ...

...low, redesign the standard curve dilution series with lower dilution factors. If the unknown sample’s ... smaller dilutions.        As mentioned above, it is a good idea to run the standard dilution series in...every run.  When setting up your dilution series, use consistent dilution factors, change pipette tips regularly...sample. For example, a BCA assay uses a serial dilution of a control protein, incubated alongside an unknown...curve? You create a standard curve by serially diluting a known control sample, called a standard. The...The ideal standard curve has at least five dilutions, with each step of the series diluted by the same ...factor. For example, Figure 1 depicts a 2-fold dilution series. Each step of the series dilutes by 1:2...

... start with a mild stripping solution and move onto a stringent solution if the mild is not effective.... pre-made stripping solutions or kits. Mild stripping The mild stripping solution uses low pH to disrupt... For the mildest stripping solution, omit the Tween 20. Once your solution is made, you’ll want to: Rinse...identify a protein of interest in a complicated solution like cell lysate. But they can be a lot of work...In this case, we recommend using mild stripping solution (see below) to minimize loss of your sample from...nitrocellulose membranes are brittle, so proceed with caution. Unlike PVDF, they can be returned directly to ...effective. You may choose to make your own stripping solution using a recipe like the ones shared by Bio-Techne...

...the glass/water interface. Super resolution microscopy Super Resolution Microscopy allows for imaging below...contrast (i.e. optical sectioning (2) or super-resolution (3). Fluorescence microscopy techniques   Fluorescent...Sheet Microscopy Light Sheet Microscopy typically utilizes a configuration of two or more objectives to create... below the diffraction (resolution) limit of a light microscope. Due to the wave nature of light, an infinitely...techniques provide a 2-10x improvement in the resolution of a light microscope (see Figure 1). What microscopy...movements. Depending on the sample, a temporal resolution on the millisecond scale may be needed. Most ...technique can be used to provide high 3D spatial resolution while also avoiding the toxic effects of high...

...prediction of therapeutic proteins. European journal of pharmaceutics and biopharmaceutics : official journal...Monoclonal antibody drugs are popular therapeutics for a plethora of disease conditions, from cancer ...response. T cell epitopes are found in either the therapeutic antibody itself or as impurities in the formulation... algorithm to evaluate the immunogenicity of therapeutics in silico. These algorithms are trained on vast...in silico assessments, as well as analyze the contributing factors that in silico prediction algorithms...algorithms fail to capture. In vitro evaluation of ‌therapeutic immunogenicity is typically performed using traditional...In-vitro models for immunogenicity prediction of therapeutic antibodies. Used under CC BY fromGroell et.al...

... Types of immunotherapeutics In lieu of the cancer-immune system interactions, therapeutics have been ...therapies include radiation therapy (owing to contributions from Madame Curie), chemotherapy, and surgical...boost the activity of the immune system, the therapeutic ability of even these conventional antitumor ...proteins are one of the most commonly used immunotherapeutic agents. Currently the FDA has 12 approved ...tumors. Attempts have been made to use whole cell therapeutics consisting of various types of immune cells....cells (DCs) are particularly popular whole cell therapeutics because they can help start a sequence of immune... cells have also been tested as vaccines and therapeutics. Their uptake by DCs is increased and was shown...

...tools, to more scientific field-based solutions or open source solutions. In comparison to paper-based record...that, academic institutions tend to think about the long term, so they need a solution for which they ...organizing folders in complex ways, there are solutions that bring more to the table in terms of data ...understand the situation in depth and provide the solution that fits the most labs, our team conducted a ... once they establish the system within their institution. In the study, respondents were also asked about...comment reflected the desire for a flexible generic solution, rather than an ELN designed for a specific research... in the near future? As most payable software solutions have user-based pricing, it is good to know how...

...research tools: Sleeping beauty, PiggyBac, and Tol2.  Sleeping beauty Sleeping Beauty is a synthetic transposable...insertions can be detrimental, it also drives genomic evolution and is used as a tool by scientists to move DNA...fish (Sandoval-Villegas et al., 2021). Sleeping Beauty’s preferred target site for integration is TA dinucleotides... efficiency decreases with cargo size. Sleeping Beauty has a close-to-random integration profile in mammalian...transposition activity (Voigt et al., 2016). Sleeping Beauty plasmids  piggyBac Although its name suggests...and DNaseI hypersensitivity sites. Like Sleeping Beauty, piggyBac has an integration efficiency in human...caused albinism (Koga et al., 1996). Unlike Sleeping Beauty and piggyBac, Tol2 has a weak consensus sequence...

...-Assisted Continuous Evolution (PACE) and Phage-Assisted Non-Continuous Evolution (PANCE) selection systems...rate of dilution (passage from the infected to a fresh uninfected host cell culture). This dilution step ...overall dilution and less stringent phage selection. Figure 2: Phage-assisted evolution of base-editing...certain sites, or make only a subset of base substitutions.  The need for speed: base editing before Cas...These systems allow for rapid continuous protein evolution through many rounds of mutations, selection, and...followed by higher stringency PACE, resulted in the evolution of TadA variants with much higher base editing... engineer and optimize base editors? Targeted evolution of naturally occurring enzymes and functional ...

...cap: institutions of higher education nonprofit entities related to or affiliated with institutions of ...role at a prestigious institution: This criterion is consistently under scrutiny. USCIS wants you to prove... Trump signed a "Buy American, Hire American" Executive Order which sets broad policy intentions directing...exempt from the cap if the applicant works at institutions of higher education, a related or affiliated...of the work of those in your field. Original contributions to the field that have had a major impact. For...and your continued work is necessary to create therapeutics to cure the cancer. Authorship:  Publishing ...professors. Casseus Law has seen that research institutions are willing to sponsor postdoctoral researchers...

...developed to break through the resolution barrier while still utilizing fluorescence microscopy, is another...previously only visible via techniques like super-resolution microscopy or electron microscopy. Since its ...light microscopes are limited in the level of resolution that they can achieve. This means that regardless...Dani, et al., 2010). How do you overcome this resolution limit? Electron Microscopy (EM) is one option...specific targets you can label in a sample. Super-resolution microscopy (SRM), a family of imaging techniques...more accessible imaging equipment, ExM protocols utilize relatively inexpensive reagents and are readily... What are some of the limitations?  A “super-resolution” technique that uses common reagents, straightforward...

...three tabs.  Each of the three tabs - Contributions, Indel Distribution and Traces - provides particular details...analysis screen. ICE analysis details Contributions   The Contributions tab shows the inferred sequences present...the most 5’ cut site.     Indel Distribution In the Indel Distribution tab, you’ll find an Indel plot ... blog. CRISPR-based genome engineering has revolutionized the gene editing field by making experimental... useful measure to determine how many of the contributing indels are likely to result in a functional ... interested in understanding how many of the contributing indels are likely to result in a functional ...plot (left panel) which displays the inferred distribution of indel sizes in the entire edited population...

...diagnostic tools and therapeutics. In addition, the strategic use of whole phages as therapeutics, which is also...blocking solution (around 108 CFU/mL is a good place to start) and add your membranes to the solution, shaking...little about how they infect and influence the evolution of their bacterial prey. Currently, receptor binding...be understood. Therefore, in addition to their utility in the health sector, phages represent a reservoir...overnight at 30°C is a good start. Prepare a lysis solution containing your favourite bacterial protein extraction...thick filter paper and saturate each with lysis solution (~3 ml each), pouring off any excess. When your...membranes off the plates and place each onto a lysis solution-soaked filter paper (colony-side up) and leave...

... surface ADRB2R, ADRBR, B2AR, BAR, BETA2AR AGRP agouti related protein homolog (mouse) AGRT, ART, ASIP2...

...targeted point mutations for directed evolution Directed evolution is an important method for protein engineering...specific agonists for orphan GPCRs demonstrating the utility of this platform for drug discovery. Researchers...Ras oncogenes have been difficult to target therapeutically due in no small part to the central role Ras...CCP); the latter of which greatly enhances the utility of this collection as it can be used to easily ...CRISPR library Addgene has had the privilege of distributing several human and murine CRISPR pooled libraries... allows for easy assembly of DNA fragments by utilizing regions of sequence homology (usually 30-40bp ... are available at Addgene. Since GMAP assembly utilizes 5 different overlap sites, this technique also...

...all-in-one acid-guanidinium-phenol solution that combines the homogenization solution and phenol addition of the...and less of being left with a bunch of random solutions when all the spin columns run out. In this article...suppliers and glassware should be treated with a DEPC solution for 1 hour, and autoclaved to remove residual ...homogenized spleens in a guanidinium thiocyanate solution and spun down the homogenate to remove the insoluble...homogenized and lysed in a guanidinium thiocyanate solution. However, instead of RNA separation using cesium-chloride...RNAProtect® from Qiagen, are ammonium sulfate-based solutions that work by inhibiting RNase activity within ...use of TRIzol® and ammonium sulfate stabilizing solutions can be made in house.   A common problem with ...

...repository has grown to include many CRISPR tools.  Distributing and depositing In Figure 1, you can see cumulative...cumulative numbers of CRISPR plasmid distribution and deposits (Figures 1A and 1B); the cumulative number...each year.    Figure 1: Addgene CRISPR distribution and deposits. (A) Cumulative CRISPR plasmids...Figure 2, the numbers on the X axis refer to distribution and deposits per year, rather than a cumulative...   Figure 2: Addgene CRISPR deposit and distribution by function and expression type. (A) Cumulative...4).    Figure 4: Addgene deposits and distribution by country. (A) Total CRISPR plasmids deposited...publication, “The Expanding Dissemination and Distribution Patterns of Diverse CRISPR Plasmids by Addgene...