We narrowed to 1,045 results for: ins;
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TypeBlog Post...difluoride (PVDF) membrane, which is sturdier and retains the blotted protein better than a nitrocellulose...methods. If you are planning to detect multiple proteins through several rounds of stripping and reprobing...membranes recommend re-wetting with methanol before rinsing with wash buffer, as dry PVDF won’t evenly re-absorb... buffer. Be sure to check the manufacturer’s instructions for the membrane you are using. Dry nitrocellulose...20. Once your solution is made, you’ll want to: Rinse the membrane in water or fresh buffer. Cover the...bad!), so be sure to use it under a fume hood. Rinse the membrane in water or fresh buffer. Cover the...imaging step to check if any secondary antibody remains, whether that involves adding fresh chemiluminescent...
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A Look Back at One Year of Plasmid Sharing for COVID-19 Research
TypeBlog Post...and identified interactions of these proteins with human proteins. Their deposited plasmids expressed ...over 75 countries. Figure 1: SARS-CoV-2 proteins and other components. Image from Maya Kostman ...infection and how to interfere with it in a BSL-2 lab instead of a BSL-3 lab. Jesse Bloom’s lab published a protocol...receptor for SARS-CoV. The lab also found that cathepsins play a role in SARS-CoV infection and deposited...Biering et al., 2021). In their search, they found mucins have an important role in viral entry, which are...viral infection. Expression of SARS-CoV-2 viral proteins In the early weeks of the COVID-19 pandemic, the...expressed 26 of the 29 SARS-CoV-2 proteins which all but one are cloned into pLVX vectors (for lentiviral packaging... -
Hot Plasmids and Viral Preps - March 2021
TypeBlog Post...Epigenetic engineering with single-pot assembly By Eric Perkins Though it’s easy enough to contemplate DNA in terms... histone-modifying enzymes and reader-effector proteins. b) Golden Gate assembly (backbones available ... create multiple synthetic chromatin-modifying proteins quickly and accurately. Priode and Haynes, ... is a new tool developed by Rui Tang in Monte Winslow’s lab to label cells after cell-cell interactions...C. elegans since 2015 to conditionally degrade proteins of interest in cells (Zhang et al., 2015). Recently.... TIR1 interacts with endogenous Skp1 and Cul3 proteins to form a SCF E3 ubiquitin ligase complex. In ...protein degradation the Ward lab used fluorescent proteins (FPs) fused to the AID* (minimal 44 amino acids... -
Cpf1 Update: Comparison to Cas9 and NgAgo
TypeBlog Post... lab characterized two Cpf1 nucleases, distant cousins of well-known Cas9. Cpf1 cleaves DNA in a staggered...indicated below each graph. Figure adapted from Kleinstiver et al., 2016. Until recently, two major elements...off-target editing efficiency. Kim et al. and Kleinstiver et al. characterized genome-wide editing efficiency... to single mismatches, but variably so, with Kleinstiver et al. reporting that Cpf1 can tolerate mismatches...essential function in Cpf1-mediated editing, as Kleinstiver et al. saw no decrease in Cpf1 activity with ...activity than that previously observed with SpCas9. Kleinstiver et al. suggest that AsCpf1’s off-target rate ...eSpCas9 and SpCas9-HF1. Both AsCpf1 and LbCpf1 ribonucleoproteins (RNPs) failed to induce off-target editing... -
Congratulations, Deck The Lab winners!
TypeBlog Post...AnltWfKuQg pic.twitter.com/OOrwo9feyJ — BTC Institute (@BTCInstitute1) December 23, 2020 Deck the Lab: ...no other, with many scientists working remotely instead of from the lab. Nonetheless, Addgene’s annual ...showcase it all! View this post on Instagram A post shared...TarlingVallim) December 19, 2020 In third place, the BTC Institute decorated with dozens of individual virus snowflakes... to #DeckTheLab with virus snowflakes from @CVRHutchinson #SciArt #STEAM #WinterWonderlab @Addgene Make...Congratulations to the winners! We’ll DM you with instructions to claim your prize. And thank you to everyone... Aliyah Weinstein... -
9 tips for a successful postdoctoral experience
TypeBlog Post... at the National Institutes of Health, was an Associate Professor at Albert Einstein College of Medicine...Hughes Medical Institute. His interests include the quality control of secretory proteins in the Endoplasmic... Research Campus of the Howard Hughes Medical Institute. Today's postdoctoral fellows (PFs) face a number...fellowship lasts five years. Most universities and institutes annually advertise for Group Leader positions...new pieces of data, your talk will probably not inspire you or the audience. Ask yourself: What is the...because the search committee asked someone at my institute about me, even though I had not asked this person...successes can be infrequent and a hobby that provides instant gratification can help boost your ego in between... -
Antibodies 101: Validation
TypeBlog Post...difficult to distinguish between proteins bound by the antibody vs proteins bound to the target protein, ...to confirm that your antibody is not binding to proteins outside of your target protein. Show suitability...antibody can distinguish between two very similar proteins. Additionally, they require variable expression...These strategies use multiple, unique antibodies against the same protein, each targeting a different epitope...correlation between the results. Using antibodies against different epitopes on the same target reduces the...This approach uses immunoprecipitation to capture proteins from a sample using the antibody and then uses...uses mass spectrometry to identify the captured proteins. If the antibody is specific, then you expect that... -
Plasmids 101: 5 factors to help you choose the right cloning method
TypeBlog Post.... Multiple inserts: There are some cloning methods that make working with multiple inserts a breeze. Among...large number of inserts to assemble. You could also consider using Gateway if your insert number is a bit...another important consideration is: The number of inserts ...as some methods are more amenable to combining...Here you simply PCR-amplify your vector and your insert, DpnI digest away the templates, and transform ...toposisomerase I, and a taq polymerase-generated insert. When mixed, topoisomerase I provides both the ...enzyme mixes. After generating the appropriate inserts by PCR, there is no need for restriction digests... very high chance of success (>95% for a single insert), mostly due to the use of toxic ccdB to prevent... -
Hot Plasmids: Winter 2025
TypeBlog Post...useful not only for future studies of kinesins but for many other proteins of interest too! Find CLIP, CLAP...brighter and more photostable than fluorescent proteins, making them ideal for single-molecule fluorescence...FRET to study the intramolecular dynamics of the proteins in solution and when associated with microtubules...microtubules. Figure 2: CLIP-SNAP fusion proteins for single-molecule FRET. A) Schematic of a CLIP-SNAP...flexible linker where a protein of interest can be inserted. B) Protein expression and labeling procedure....deposited an array of constructs for tagging the kinesins KIF5A and KIF5B as well as backbones for generating...generating other SNAPf-, CLIPf-, and CLAP-tagged proteins for mammalian cell expression. These plasmids will... -
Experimenting with New Careers while in Grad School
TypeBlog Post...experience with my blog helped immensely when CINSO started brainstorming the resources we wanted to provide online...and other nonprofits to gain support for CINSO. For instance, I tweeted to the Indiana State Museum and...nonprofit called Central Indiana Science Outreach, or CINSO. We organize fun science events for adults and professional...called “nontraditional,” helped prepare me to start CINSO. Here, I share some of the tips I learned along ... best supporter and most productive partnership CINSO has had. Volunteer! The turning point in my graduate... and graduate school resources daily to promote CINSO events and recruit speakers and participants for...something I had zero experience with before founding CINSO. Now, the network I built during graduate school...