... are not convinced by the existing data, you may need to roll up your sleeves and perform some extra validation...that it will perform as expected in another. You’ll need to validate that points 1 and 2 hold true under ...different tissues or cell types. If your antibody indeed binds to your target then you would expect to see...expression and non-specific, background signal, so you’ll need a range of expression strengths to be confident ...unexpected. This approach has high specificity, but you need to be aware of any possible alternative translation...identified as your target protein, allowing you to proceed with confidence.  Share your results Ultimately...

...mastery. Scientists need to be doers for a long time before they master the skills they need to be successful...these three roles allows you to identify what you need to do to be successful in a specific role or cycle...hands-on bench work. To really be happy, you may need to find a way to do, manage and lead all in one ... opportunity to take on the role of leader, one needs to take the initiative by bringing and effecting...

...cloning process - please check out that post if you need a more detailed refresher on the downstream steps...restriction site to the reverse primer. Next, we need to examine the DNA sequence that we want to amplify...the Reverse Primer, the design is similar, but we need to use the reverse complement to get PCR amplification... (30bp with 18bp of homology to the ORF). We now need to generate the reverse-complement of this sequence...wide range of annealing temperatures, but you may need to increase your primer length and increase the ...

...fertilizer, seeds and a growth space with fluorescent lights. Other experiments require only seeds, petri ... to collect, preserve, reproduce and distribute seeds, DNA and other resources for Arabidopsis and related... free-of-charge from the ABRC Outreach website. Seeds, which are free for K-12 teachers and available ...require a huge investment in resources. The materials needed to conduct the experiments outlined in ABRC’s kits...

... be a huge challenge. Epitope tags eliminate the need for target-specific antibodies and have been widely...fortunate enough to study actin, then you may not need to read this - but for many other proteins, quality...tracking your protein may also be possible, without the need for an antibody. Tags can enable the separation ... Myc The Myc tag is a 10 amino acid peptide (EQKLISEEDL) derived from the protooncogene of the same name...interfere with protein function and localization. If you need monoclonal GFP antibodies or guidance on designing...

...Initiative, will allow Addgene to create a much needed open-access recombinant antibody/affinity reagent...produced in animals may be variable among animals and bleed dates. The development of monoclonal antibody (mAb...neuroscience research today, but there is a critical unmet need for an open-access, molecularly defined, and well...Trimmer. But in order to create such a resource, we need your help. You can participate by depositing your...share, edit, and access the affinity reagents they need to make their next big discoveries. We welcome you...

...easy to find in publications, so this is a much-needed community resource. Instead of painstakingly searching...use the AAV Data Hub to quickly find details they need for planning their next experiments using a specific...image yourself and it is under copyright, you will need to obtain written approval from the copyright owner... that this provides a clear overview of what you need to submit data. Please note that the data hub is...after submitting or viewing the data you have any feedback at all, we’d appreciate you sharing it through...

... bottom line is that a lot of AAV particles are needed to get a few good ones into a cell to produce expression...transport and transgene expression It’s clear that we need high titers, as contact between the cell and any...protein. However, expression of receptors may not need to be as high to achieve an observable signal in...in your experiment, therefore titer may not need to be as high when delivering receptor constructs with...improve AAV production.     References 1.Clark, K. Reed. "Recent advances in recombinant adeno-associated...

...also use these tips as a starting point when you need to amplify a library for your own experiments. ...individual plasmid. However, lots of library DNA is needed for screening experiments, so the scale of the ...for most libraries. Here are the key points you’ll need to remember when undertaking library amplification... are identical except for the insert; there’s no need to sequence other parts of the plasmid. How will...reads is, again, seemingly straightforward – you need to count how many times each of the library components...

... studying. You need to make sure your assay is scalable and robust. What you don't need to do is spend... is to quickly generate a single cell line and proceed to use it for experiments.” - Alex Chavez, researcher...available as lentivirus and we’ll be adding more as the need arises. For example, we’ve teamed up with the Broad...When you're setting up to do a genetic screen, you need to focus on building a really good model that reflects...

...will also need to be perfected until the efficiency of repair approaches 100 percent. “We need to be sure...currently being tested in clinical trials. It is speeding the process of the discovery/development of genetic...advantage over induced pluripotent stem cells, which need to go through a phase of crisis where they 'collect...effects are important,” she said. “We also really need to get a handle on how cleavage sites are repaired...

...educational content Addgene truly believes that sharing speeds science, and our entire mission is to enable that...our frequently asked questions!    Identify needed content Let’s take a look at how we create this...first step was to understand information our guide needed to have. Through a series of conversations on resources...basic information someone new to antibodies might need to successfully start using them at the bench, along...in which they have expertise. Other times, we may need to reach out to an external expert to help us create...

...thousand plasmids from Ginkgo. To balance the need for speed while ensuring our rigorous QC standards are...the reach of these plasmids is indeed global. Helping scientists speed up research is what we’re here ...To get these plasmids available for scientists speedily meant that we had to revamp some of our quality...

...provide a much needed boost to basic molecular plant research, more initiatives are needed to increase the...and opportunities abound to build better fuels, feeds, foods, and fibers. And because working with plants...that the scientific and the agricultural community needs to reclaim the term “GM” by demonstrating the current...the day cannot be developed through conventional breeding; synthetic biology is required to achieve this...

...Researchers need to learn sophisticated methods of sex and gender analysis. Universities need to incorporate...research that fails less and helps more people, we need to take sex into account. Gendered Innovations is...Especially in animal research, they worry about needing more study subjects and control groups. The important...these methods into their curricula. Corporations need to integrate these insights into product design....

.... 2. Studying negative feedback loops of signaling pathways Negative feedback loops are a common feature...kinase activity. Zhou et al studied the negative feedback on endogenous MEK1/2 phosphorylation by creating...PP1/PP2A is responsible for this MEK1/2 negative feedback. 3. Studying signaling pathways in vivo Expressing...possibly be used in other model systems, but they need to be expressed in a tissue or organism that blue...

...and they provide different amounts of access or feedback. One of the most significant shifts in scientific...of strategies for your scientific communication needs. Going to present some unpublished results at a ...preprints as part of their scientific communication needs. The most often-cited reasons researchers might ... in most review processes. Preprints and their feedback, in contrast, are immediately and openly accessible...formal peer review, with the earlier attention and feedback giving authors the opportunity to strengthen the...

...secondary should be used when signal amplification is needed.  Species  Next, check the species that the antibody...a good chance that the antibody could meet your needs.    Multiplex experiments Finally, consider whether... antibodies in a single experiment then you will need to make sure that you can easily distinguish each...purchase but also someone to turn to for advice if you need to troubleshoot. What do you know about the vendor...validation does not mean that the antibody is guaranteed to work. Antibody-based assays are complex and...

...technique, and sometimes each individual assay, will need to be developed and optimized for the samples, antibodies...multiple proteins of interest - the secondary antibody needs careful consideration before selection. If all the...precious commodity in the lab and eliminating the need for secondary antibodies would save a good deal ... antibody labelled with a probe, eliminating the need for a secondary antibody. Other immunohistochemistry...option for signal amplification while providing needed flexibility in assay development. Researchers who...

...interpretations of what we discussed and decided. I needed to a) figure out the sources of miscommunication...almost certainly experienced some of the hurdles I needed to overcome.  I eventually pinpointed and (mostly... software engineers in your lab, you likely will need to write protocols, SOPs, and methods sections. ... not have the context you do, which means you’ll need to provide that context yourself. Simple words ... commonly used (and misused) words that clearly needed an Addgene-specific definition. Words such as “...