...because it turns a dynamic process into a static measurement and it provides limited information about what...fluorescent autophagy reporters now allows for the measurement of autophagic flux, or the changes in autophagic...strength excited at 586 nm to 440 nm serves as a measure of autophagy induction: when Keima is in the cytosol...is high. When untargeted, Keima can be used to measure bulk autophagy, but the addition of a mitochondria-targeting...3B, or LC3. LC3 is a commonly used marker for measuring autophagic flux. mRFP-GFP-LC3 functions as an ...18094608. Sargsyan, A., et al. "Rapid parallel measurements of macroautophagy and mitophagy in mammalian...Saori R., and Noboru Mizushima. "Monitoring and measuring autophagy." International journal of molecular...

...organic solvents’ impact on lipids means that they are able to permeabilize your sample while fixing it, which...Examples of different permeabilization strategies and when to use them. A) Not permeabilizing is suitable for... first couple of steps - mainly, fixing and permeabilizing. Fixation What is the point?  Fixation allows...the protein. Created with BioRender.   Permeabilization What is the point?  Antibodies are large proteins...they are not able to diffuse across natively. Permeabilization essentially punches holes in the cell membranes...In fact, you may get cleaner results by not permeabilizing as you’ll prevent binding to any target protein...of the cell. However, assuming you do need to permeabilize, you have a few options for what reagents to...

...viral vector solution, and generally these methods measure different characteristics of the solution. These... containing viral genomes. Physical titers are measured by quantifying the concentration of viral genomes... it is important to understand what these values mean and how they can be compared.   Figure 2:...circled in red) report physical titer, which is measured by qPCR.   Physical titer is typically calculated...repeating the exact same PCR twice can give titer measurements that vary up to two-fold. How can we get more...Taylor et al., 2017). What do Addgene AAV titers mean for your experiments? Because (physical) AAV titers...

...important for regulating signal transduction and measuring kinase activity at the single-cell level can aid...whose cellular localization serves as a proxy measurement of kinase activity. The key advantage of KTRs...Advantages of KTRs When compared to FRET reporters for measuring kinase activity, KTRs have two key advantages....nucleus, while FRET-based reporters can be used to measure kinase activity in specific subcellular compartments... Regot, S., & Covert, M.W. (2018). Live-cell measurements of kinase activity in single cells using translocation...

...Quantitative measurements of membrane charges A FRET sensor enables quantitative measurements of membrane...of plasmids encoding fluorescent biosensors to measure biomolecules or genes via FRET or other assays....enable investigation of a signaling pathway or measurement of a biomolecule while largely preserving spatial...ratiometric calcium sensor enables quantitative measurement of the local calcium microdomain in the endoplasmic...sensors ZapCY1 and ZapCY2, Golgi or ER targeted Measuring steady-state and dynamic endoplasmic reticulum...di-GMP FRET-based biosensor for cyclic-di-GMP measurement Asymmetrical distribution of the second messenger...Akerman Chloride (Cl-) YFP-derived sensor for measuring Cl- concentration in physiological range A genetically-encoded...

... two types of antigen specificity: monoclonal, meaning many copies of one clone that are specific to a... a certain epitope, or polyclonal, meaning there are many clones that are specific to multiple epitopes...bound. Signaling molecules , proteins that emit a measurable light or color in response to a specific stimulus...proteins. HRP is also used in ELISAs, which only measure a single antibody signal. Here, HRP is used as ...samples can be normalized to a total protein measurement. Both methods will also serve as a control for...conjugate, typically HRP, is activated and its output measured. This method can be performed using the indirect...Fix your samples. Prepare your samples using permeabilization, antigen retrieval, and/or sectioning. Block...

...energy transfer is measured using fluorescence microscopy by exciting the donor and measuring the emission ...collection of plasmids suitable for FRET experiments to measure protein interactions or conformation changes, as...calibrate the numerous and disparate methods used to measure FRET. Plasmid FRET Pair C5V Cerulean attached to...

...fluorescent protein variant (CyPet) optimized for FRET measurements with a yellow fluorescent protein variant (YPet...fluorescent protein variant (YPet) optimized for FRET measurements with a cyan fluorescent protein variant (CyPet...around a bacterium. Plasmids containing easily measurable reporter genes (eg. LacZ or Fluorescent Proteins...it. Bacterial two-hybrid systems can be used to measure protein-protein interactions; if two proteins of...

...with Coomassie blue. Protein band intensities are measured using ImageJ software and a standard curve is ...intensities of both the antibody and impurities are measured and the ratio of the antibody protein content ...Immunohistochemistry (IHC) Tissue sections are fixed, permeabilized, and labeled with the antibody and a species-specific...

...amplification, and high-quality quality control measures such as titering, purity assessment, sterility...institution’s approval of the MTA, and shipping, mean that the actual time from request to delivery could...including purification, titration, and quality control measures. We have distributed preps around the globe and...

...based on the gRNA sequence without cleavage. This means proteins with other functions can be fused to a ...Cas9s (hfCas9) . There is currently no defined measure of fidelity, so sometimes they will be referred...principle, the ability to induce base transversions means that any base-to-base conversion is possible with...CASTs can result in bidirectional insertions — meaning they may not be suitable for applications where...Partridge, E. C., Newberry, K. M., Smith, S. B., Meadows, S. K., Roberts, B. S., Mackiewicz, M., Mendenhall...

...protein, and is suitable for both cell-permeant and non-cell-permeant proteins. CLIP-tag CLIP-tag is identical...

...your gene "in frame" with the fusion protein. This means that the final product should be translated...Representative Empty Backbones Promoter Measure promoter strength pBV-Luc - Luciferase ...

... must be delivered intracellularly in order to measure luminescence. Gaussia luciferase is secreted and...reporter. Unless otherwise noted, these plasmids are meant for expression in mammalian systems. ID Plasmid ...

...application-specific validation and consistent quality control measures. We work with trusted partners and experts in ...

...Optogenetics Plasmids | Guide Use light to detect, measure, and control molecular signals, cells, or groups...

...as microbial opsin and optical switch tools to measure and manipulate cells and their biomolecular processes...

...concentration given relatively to the non‐targeting spacer measured by high‐throughput microscopy for a collection...

...examples of reporter plasmids that can be used for measuring infectivity and neutralizing activity are listed...

...function of any viral vector. Addgene performs QC measures which verify the identity of the viral vector,...