...proteins tagged with genes or peptides with known subcellular localization patterns. Useful for co-localization...Proteins Organelle Markers Fluorescent Proteins: Subcellular Localization More Fluorescent Protein Resources...Cytoskeletal Junctions Secretory Pathway Background The subcellular localization of a protein is often tied to its...There are a variety of methods for determining subcellular localization. One of the most common is to use...proteins that are targeted to various organelles or subcellular structures. You should fuse your protein of interest...The cell contains a variety of organelles and subcellular structures each with their own unique function...

...pDONR221). These vectors allow a fast and efficient sub-cloning into a wide range of Gateway-compatible pDEST...

...and AscI at the 3' end. These inserts can then be sub-cloned into a series of expression vectors which ...

...enzymatic binding to attach substrate Control plasmids with a recognizable sub-cellular localization (membrane...information on substrates and synthases needed for use of these tags. Plasmids Addgene ID Plasmid Substrate Feature...simultaneously with dual labeling when used with two substrates Can be used in live or fixed cells, and for internal...protein tags based on the acyl carrier protein. Substrates are derivatives of Coenzyme A (CoA). These tags...252 KB) ACP and MCP tags ACP-MCP-Tag plasmids - Substrate source (PDF, 46 KB) Please note: New England Biolabs...product portfolio, including the CoA-derivative substrates and the ACP and SFP Synthases. Users may still... their own fluorescent or non-fluorescent CoA substrates by purchasing coenzyme A disodium salt from a...

... expressing these tools in well-defined cellular sub-populations has generally been limited to single-...recombinase-expression strategies to define cellular sub-populations of interest, including dual-transgenic...opens in a new window) Hafner G, Witte M, Guy J, Subhashini N, Fenno LE, Ramakrishnan C, Kim YS, Deisseroth...projections of molecularly defined dopamine neuron subtypes using intersectional genetic approaches. Nat Neurosci...

... out Joanne's Reddit AMA Here are 6 steps to submitting a resume that will give you a chance at an interview...interview:  1) Have an excellent resume ready to be submitted The structure and content of a resume can be crucial...the hiring group.  Read here for more on timely submission of a job application. 3) Read job descriptions... not likely know the exact person monitoring submissions (usually a Human Resources representative). Pro...

...and selected subcellular marker proteins, it’s possible to get an idea about the subcellular structures ...protein function. Getting ready to determine the subcellular localization of your protein of interest The ... vesicles via the secretory pathway to their subcellular or extracellular destination. At the end of their...proteins tagged with FPs to highlight various subcellular structures. Please see Figure 1 for commonly ...inert fluorescent protein fusions to visualize subcellular structures. For example, you might use your fluoresent...particular gene. For subcellular markers in yeast see our plasmid collection from...You can find more plasmids for labeling your subcellular structure of interest in the Allen Institute ...

...Plasmid 1: B. subtilis knock out and knock down libraries. Listen to Our segment on the B. subtilis libraries...you'll learn about new libraries for studying B. subtilis biology, plasmids for enhancing iPSC production... b. subtilis...

...where your plasmid yield is sub-optimal? If you are consistently getting sub-optimal plasmid yields from...your stab from Addgene, streak for single colonies, sub-culture, and prep with a DNA prep kit or your lab's...plasmid yield. Start with a single, fresh colony Sub-culturing directly from a frozen glycerol stock or...

...Visualization of neuronal sub-cellular structures GFP is limited in its ability to label subcellular structures in...be used to amplify the direct smFP signal within sub-cellular structures. For instance, smFPs have been... be very effective probes for finely visualising sub-cellular structures or low-abundance proteins. When...

...of local temperature sensing with sub-millisecond resolution and sub-degree temperature differences under...temperature variations and heat transfer at the subcellular level requires a probe with fast kinetics and...

...Chylinski (who coincidentally speak the same Polish sub-dialect making their Skype communications extra fun...

...repair template strategies activate different HDR sub-pathways like synthesis-dependent strand annealing...DNA repair (with bonus gRNA design resources.) Subscribe to CRISPR updates on our blog to keep current ...

...provide a true measurement of high affinity (i.e. sub-nanomolar) protein-ligand interactions. Avidin and... calculate the amount of bound ligand (after subtracting out non-specific binding). Historically, glutathione-S-transferase...to express a foreign protein in bacteria and subsequently purify the fusion protein from a bacterial extract...extracellular domain of the erythropoietin receptor and subsequently evaluated interactions with mutant and wild-...

...regular group meetings, journal clubs, and focused sub-group meetings to discuss day-to-day progress, ideas...

...direct your biosensor to specific tissues or cell (sub)types for expression and downstream monitoring. Nervous...

...For example, calcium in the ER lumen reaches the sub-millimolar range, much higher than in the cytosol...population of cells (e.g., a neuron subtype) and specific subcellular compartments for measurements. All...Localization The calcium concentrations in different subcellular compartments can vary greatly. Additionally, ...changes you will observe in your experiment could be subtle or dramatic. You want to use a calcium sensor that...allosteric rearrangements of the sensor, and (3) subsequent change in fluorescence only occurs at high calcium...sensor to specific tissues or even distinct cell subtypes for efficient expression and downstream monitoring...

...the desired edit to still occur if the cut site is sub-optimal? As with everything in biology, the answer...little flexibility in the location of the cut and subsequent edit. In these applications, location takes priority...requirement to allow flexibility in gRNA selection and subsequent editing. Identifying PAM-free and -flexible Cas...

... genome-wide CRISPR libraries and a selection of sub-libraries targeting specific classes of human genes...

...of stochastic magic and Poisson statistics, each sub-population has, on average, one distinct enhancer...