We narrowed to 786 results for: KIN
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TypeBlog PostPublishedJuly 10, 2014, 7:17 p.m.... and devoted 100% of his energy and effort into making the startup self-sustainable. Within a year from...all safety nets.” Raising seed money. “Before seeking funding, ask yourself if you really want to do ...Co-founder and formerly VP Operations of Sample6) for taking the time to speak with us about the biotech startup...
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Antibodies 101: Affinity Reagents
TypeBlog PostPublishedApril 14, 2022, 1:15 p.m....someone speaks about an affinity reagent, they’re talking about a standard antibody: An ~150kD protein with...nanobodies, are derived from a camelid antibody lacking a light chain, called a heavy chain antibody (Muyldermans...clear: it’s an exciting time to be a scientist looking for that perfect fit! References and Resources... -
Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options
TypeBlog PostPublishedMay 11, 2021, 1:15 p.m....between them, could affect the properties or trafficking of the fusion protein (Kimple et al., 2013). ...opsin-expressing cells). For other things to keep in mind when making fluorescent protein fusions, check out Joachim ...Fluorescent Proteins 101: GFP Fusion Proteins – Making the Right Connection. Thank you to our guest... -
Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins
TypeBlog PostPublishedMarch 19, 2019, 1:31 p.m....inhibition is spatially and temporally ubiquitous, making them a rather blunt instrument for controlling ...and systematically mutated it. Using a robotic picking approach to select cells with variant characteristics...al, Nat Chem Biol 2018. PubMed PMID:29483642. Making protein-protein interaction studies easier with... -
Antibodies 101: Fab Fragments
TypeBlog PostPublishedSept. 5, 2024, 1:15 p.m....this point, you may be asking yourself, what type of Fab fragment are we talking about here? Many of the...fragment. But this bivalency may cause problems for blocking purposes, since the fragment could bind not only... -
Choosing the B(right)est Fluorescent Protein: Aggregation Tendency
TypeBlog PostPublishedJune 15, 2017, 2:30 p.m....in some applications (e.g. labeling of cells or tracking promotor activity), but it is problematic in applications...for individual cells, and the number of 'normal' looking cells can be counted. This assay will provide information...on the dimerization tendency in living cells. Strikingly, the OSER assay reveals that tagRFP does not ... -
Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation
TypeBlog PostPublishedDec. 14, 2021, 2:15 p.m....laboratory of Dr. Moshe Szyf at McGill University, working to develop CRISPR-based tools for epigenetic editing...Charlton J, Mattei AL et al (2018). Genome-wide tracking of dCas9-methyltransferase footprints. Nat Commun...Zhong C, Eason DE, Ross DL, Vachhani RV, Ming G-L, King JR, Song H, Sweatt JD (2013) TET1 Controls CNS 5... -
Adenoviral Vector Production and Troubleshooting
TypeBlog PostPublishedJune 14, 2018, 11:44 a.m....density: particle concentration is measured by looking at absorbance at 260nm using UV/Vis spectrophotometry.... The purity of the sample can be estimated by looking at the ration of DNA (260nm) versus protein (280nm...calculated in terms of plaque forming units (pfu), taking initial vector dilution into account. You'... -
Fluorescent Proteins 101: Introduction to FRET
TypeBlog PostPublishedJune 27, 2017, 1:03 p.m....or the activity of a protein in a synapse. What kinds of tools enable us to measure these properties, ...articles describing the FRET tool that you are looking for and check our curated list of biosensors too... Sustarsic, Marko, and Kapanidis, Achillefs N. "Taking the ruler to the jungle: Single-molecule FRET for... -
Plasmids for Endogenous Gene Tagging in Human Cells
TypeBlog PostPublishedApril 6, 2017, 1:02 p.m....alleninstitute.org. A classic challenge in cell biology is making sure that what we observe through the microscope...whole segment—a GFP tag and 1kb of homologous DNA flanking both sides (about 2.7kb total)—is inserted into...and destroyed by CRISPR/Cas9 during transfection. Making the double strand break and introducing the tag... -
Hot Plasmids February 2024
TypeBlog PostPublishedFeb. 20, 2024, 2:45 p.m....10.1038/s41587-023-01756-1. PMID: 37095348. Breaking up is hard to do: Monomeric StayGold variants ...with some applications. Several labs have been working to produce monomeric versions that break up the...was delivered to brain endothelial cells in mice lacking LY6A using AAV1-X1 (Chen et al., 2023); then, when... -
Hot Plasmids and Viral Preps - January 2021
TypeBlog PostPublishedJan. 5, 2021, 2:15 p.m....Scientists looking for more compact genome editors due to delivery constraints and scientists seeking a wider... prep in serotype AAV5! Get dLight1.3b here Looking for a knock-out in your experiments? AAV-flex-taCasp3... -
Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux
TypeBlog PostPublishedSept. 13, 2016, 2:30 p.m....Advancement of Science, Rush Holt, followed by a networking reception. Saturday’s workshop will be an advocacy...laid out by a 2014 National Academy of Sciences working group (National Academy of Sciences. “The Postdoctoral... Advancement of Science, Rush Holt, PhD and a networking reception. Saturday’s workshop will progress ... -
5 Tips for Troubleshooting Viral Transductions
TypeBlog PostPublishedAug. 11, 2016, 2:23 p.m.... me troubleshoot some technical obstacles when working with virus: 1. Viral vectors undergo DNA rearrangements...cells for infection If your transductions aren’t working, you can check if virus is being generated by your...template. Regardless of the type of virus you’re working with, there’s a lot that goes into virus design... -
Plasmids 101: FLEx Vectors
TypeBlog PostPublishedApril 28, 2016, 2:30 p.m....recombination events, and that the orientation of the flanking loxP sites directs the Cre recombinase to invert...placement of the two pairs of target sites. By flanking the reporter with one loxP site and one lox511...PMID: 12665802. 2. Branda, CS and Dymecki, SM. Talking about a Revolution: The Impact of Site-Specific... -
Tips for Titering Your Lentiviral Preps
TypeBlog PostPublishedMarch 15, 2016, 2:30 p.m....The day has arrived; you’ve painstakingly cared for your packaging cell line, prepped your DNA, transfected...amount of virus you actually have in your prep. Taking time to properly titer your virus will not only...original plasmid which can over-inflate results. A packing vector deficient in reverse transcriptase will ... -
7 Simple Strategies to Resolve Conflicts with Difficult Supervisors
TypeBlog PostPublishedJan. 21, 2015, 1:30 p.m.... who keep changing their minds and have trouble making decisions. In my work as a thesis coach for graduate...categories. In addition, I also learned about two other kinds of difficult supervisors: Hands-off super busy ...explain how it is interfering with your work. Avoid talking about your emotions, such as frustration or anger... -
PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs
TypeBlog PostPublishedJune 25, 2024, 1:15 p.m....in brackets and add the upstream and downstream flanking sequences to their input string (Figure 2). This...prediction models. In addition, we provide options for nicking gRNAs (used for PE3 and PE5 prime editing strategies...strategies) based on a DeepSpCas9 score ranking (Kim et al., 2019). PRIDICT saves time and resources by ... -
Viral Vectors 101: Voltage Indicators
TypeBlog PostPublishedJan. 30, 2024, 2:15 p.m....components of the indicator, which can make the kinetics of voltage sensing to signal output slightly slower...fluorescent signaler. The pro of this system is the kinetic response time from signal input to readable output...FP-rhodopsin GEVIs have been engineered to have kinetics comparable to its one component counterpart and... -
Fluorescent Tagging of Endogenous Genes with SapTrap
TypeBlog PostPublishedOct. 12, 2017, 2:26 p.m.... localization, cell structure, intracellular trafficking, and protein turnover rates. Additionally, by...fission/fusion, nuclear import, and neuronal trafficking. Although FPs have enabled discovery of many ...cellular mechanisms, there are some limitations to working with FPs. Overexpression of fluorescently tagged... -
Tips for a 1st time AAV user (by a Rookie AAV user)
TypeBlog PostPublishedOct. 23, 2018, 11:49 a.m.... I’m not going to lie, AAV transfer plasmids are kind of a pain to work with. These plasmids contain ITRs...the viral particles of the AAV serotype you’re working with are primarily found in the culture media, .... If you’re new to creating gradients, practice making the iodixanol layers before doing your first purification... -
Which Fluorescent Protein Should I Use?
TypeBlog PostPublishedMay 20, 2014, 2:06 p.m....the FP (fused to the protein of interest), thus tracking the localization of that biomolecule inside living...eel was shown to mature independently of oxygen, making suitble for use in anaerobic conditions. Maturation...fluorescent protein should I use?" And if you're looking for more fluorescence microscopy tools and techniques... -
Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy
TypeBlog PostPublishedAug. 27, 2020, 1:15 p.m....cellular functions, including ubiquitinoylation, kinase signaling, integrin signaling, and transcription...Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection. Cellular expression Whether...localization may differ from those of your BiFC construct, making it difficult to interpret your results. Unfortunately... -
Plasmids 101: Secondary Nanobody Toolbox
TypeBlog PostPublishedFeb. 27, 2018, 2:04 p.m....domain proteins, they can be expressed in bacteria, making nanobodies a recombinant and renewable anti-IgG...available nanobodies from Pleiner et al. Before picking a secondary nanobody, it’s important to review ...expression plasmids here Additional Resources Looking for monoclonal primary antibodies to use with the... -
Viral Vectors 101: The Retroviral Lifecycle
TypeBlog PostPublishedJuly 27, 2023, 1:15 p.m....does not do). Furthermore, molecular biologists working with vectors are, in most cases, interested in ... the viral integrase protein (IN) during step 3 (King et al., 2008) (Hu & Temin, 1990). At step 3, the....1016/0022-2836(73)90277-5. PubMed PMID: 4355596 King, S. R., Duggal, N. K., Ndongmo, C. B., Pacut, C.... -
Droplet Digital PCR for AAV Quantitation
TypeBlog PostPublishedJan. 16, 2020, 2:18 p.m....our samples 1:6 million to 1:25 million. After making the dilutions, they are transferred to a new plate...performing ddPCR Clean everything You must have a clean working area to titer your AAV by ddPCR. Droplet digital...generator, you should avoid pipetting too quickly when making your virus dilutions. This will reduce aerosols... -
Better Dyeing Through Chemistry & Small Molecule Fluorophores
TypeBlog PostPublishedSept. 8, 2016, 2:30 p.m....dye, mauvine, was discovered in 1856 by William Perkin. His discovery set off a flurry of activity and...labeling strategies and improved fluorophores are making chemical dyes increasingly attractive and accessible...Medical Institute. He would rather be in the lab making new fluorophores for advanced imaging modalities... -
Rett Syndrome: A History of Research and Therapeutic Outlooks
TypeBlog PostPublishedNov. 19, 2020, 2:15 p.m....speech, coordination, purposeful use of hands, and walking. They may exhibit slow head growth, develop seizures...medical journal. Decades later and after years of painstaking work trying to isolate the causative gene for...:2<147::aid-ajmg6>3.0.co;2-o Baker SA, Chen L, Wilkins AD, Yu P, Lichtarge O, Zoghbi HY (2013) An AT-Hook... -
Viral Vectors 101: Pseudotyping
TypeBlog PostPublishedApril 20, 2021, 2:30 p.m.... delivery to only a few cell types. If you’re looking for more control in the cell types your lentiviral...retrograde direction from the synapse to the cell body, making it a great tool to study neuronal connectivity....HY, Tortorici MA, Veesler D, Murphy M, Pettie D, King NP, Balazs AB, Bloom JD (2020) Protocol and Reagents... -
Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More
TypeBlog PostPublishedAug. 13, 2019, 12:58 p.m....zebrafish embryo develops outside of the mother, making them ideal for studying vertebrate embryogenesis... target mRNA inhibit its translation either by blocking initiation or splicing. (RNAi, in contrast, engages...unprecedented views into cognitive processes like decision making (Kim et al, 2017). “Brainbow” fish lines have been...