...YFP and use an affinity column for purification. After binding YFP to the column, you wash several times... use OptoNBs. You fill a column with OptoNB coated beads and wrap the column with blue LED lights. When...their target protein is altered by blue light illumination. This ability to reversibly bind and release...pathway. What’s next for OptoNBs? While OptoNBs illuminate a path to light-controlled protein binding, they...

...bind the IgG heavy chain. Running the serum over a column of resin crosslinked to protein A /G will bind ... antigen. In this case, the serum is run over a column containing the target protein, and any antibody...recognizing the target protein remains bound to the column while the rest of the proteins are eluted. Then...Then, the antibodies are dissociated from the column and from the target protein. Pre-adsorption Finally... pre-adsorb the antibodies, pass them through a column containing immobilized serum protein from other...-react with the serum proteins pass through the column, resulting in a polyclonal antibody population ...

... genome copies/mL (GC/mL)), and total injection volume. You’ll often hear that “high titer” is required... It’s the optimal number of viral particles per volume required to express the gene of interest, in the...expression system for both the titer and total injection volume.  There are a few factors to consider. First, ..., the lower the titer and vice versa. The total volume needed, however, increases with the size of the...- and as your titer changes, so may your needed volume (de Backer et al., 2010).  Second, high efficiency...Engineered AAV Capsids Improve Transduction and Volumetric Spread in the CNS. Molecular Therapy - Nucleic...

...can be measured using a technique known as Bioluminescence Resonance Energy Transfer (BRET). BRET is similar...BRET is that the energy transfer is between a luminescent donor and a fluorescent acceptor. TRUPATH uses...measures heterotrimeric G protein dissociation by bioluminescence resonance energy transfer 2, or BRET2. With... constructs. Cells are then treated with the luminescent enzyme substrate coelenterazine, followed by ...endogenous ligand or drug. In the inactive state, the luminescent chimeric protein Gα-Rluc8 and Gγ-GFP2 will be...

...plasmid per row, as outlined by the column headers. Required columns are shown in red in the spreadsheet...species can be indicated in the “Species Other” column. Relevant Mutations Indicate the amino acid... If other, indicate type in “Vector Type Other” column. For example, you may enter “donor template vector...the deposit spreadsheet showing the first eight columns and a pop-up instruction box for the “Purpose” ...

...compensate for this, researchers can plate larger volumes of the outgrowth containing the transformants onto...the lacI/lacZ cassette, and illustrates how the volume of the 1 ml outgrowth spread on each transformation...fragments assembled Golden Gate Assembly protocol* (Volume of 1mL Outgrowth plated) Correct assemblies per...Efficiency of assemblies per plate using outgrowth volumes described in Figure 3, with calculated yields from...ºC incubations or cycling.  *Assembly reaction volumes were 20 μl (1, 12 fragments) or 25 μl (24 fragments...

...fluorescent proteins, and luminescent proteins. While fluorescent proteins and luminescence have been widely used... fluorescence or luminescence, which require UV lamps, fluorometers, or luminometers, chromoprotein detection...striking visuals from fluorescent jellyfish, bioluminescent phytoplankton, and colorful corals, to name...

...determine the optimal/minimal injection titer and volume needed to achieve adequate expression in your cells...cells of interest.  Determine injection titers and volumes based on the serotype Since each serotype also ...advisable to try a few different injection titers and volumes to determine the optimal injection parameters needed...will already have a starting “optimal” injection volume and titer to begin with. Now you will mainly only... vector, a range of dilutions and/or injection volumes may need to be tested with each new vector or lot...

... dyes are now commercially available as Lumio Green and Lumio Red from Invitrogen (now ThermoFisher Scientific...Acceptor Tag R0 (Å)a Range (Å)b Applications CFP   Lumio Green Tc: CCPGCC 50 40-72 Intact Cells Cy3NTAc...Cy5NTAc 32 25-45 GFP (fluorescein is similar)   Lumio Red Tc: CCPGCC 48 38-70 Intact Cells Cy3NTAc Poly...permeabilized cells  Cy5NTAc 43 34-62 YFP   Lumio Red Tc: CCPGCC 54 42-78 Intact Cells Cy3NTAc Poly...

...vivo.  Read on to learn more about this killer illumination technique! What is a photosensitizer? First...within 30 minutes of exposure. 90 seconds of illumination were sufficient for 100% effective killing of...the dye. Embryos exposed to just 12 minutes of illumination had fewer mCer3-labeled cells in their hearts...photosensitizer protein that generates ROS when illuminated with green light. They found that KillerRed required...

...however, affected by bubbles in wells, variations in volume, or machine surge errors and allow for well-to-...revisited. Biomolecular Detection and Quantification. Volume 14. 2017, Pages 19-28, ISSN 2214-7535, https://...products in a rich web application, Bioinformatics, Volume 27, Issue 7, 1 April 2011, Pages 1019–1020, https...Reproducible Data the First Time, Trends in Biotechnology, Volume 37, Issue 7, 2019, Pages 761-774, SSN 0167-7799... to mutation detection, Nucleic Acids Research, Volume 28, Issue 19, 1 October 2000, Pages 3752–3761, ...

...the volume typically used (e.g., use 5 mL medium on a 10 cm plate). By reducing the culture volume into...resuspension of the collected particles in a smaller volume. Always remove packaging cell debris before proceeding...without ultracentrifugation, you can reduce the volume of cell culture medium on the packaging cells immediately...

...are both great options for this, depending on the volume of culture you chose to work with. You will get... construct at a time. For detailed protocols, volumes, spin speeds, recipes, and Agrobacterium strains...messy, so you want to make sure you have sufficient volume to dip all your plants. Make sure your Agrobacterium... viable. The time this will take depends on the volume of culture you are using and whether you are inoculating...

...Nickel Column: Lyse the bacteria expressing your nanobody and run the lysate over a nickel column. Nanobodies...Nanobodies will bind to the column due to their N-terminal His14-bdNEDD8 tag. After several washes, this...protease, thus releasing purified nanobodies from the column. bdNEDP1 protease can be generated using plasmid...

...chromatography is a popular choice for this. Chromatography columns containing appropriate immobilized ligands are ...buffer exchange can be performed using a desalting column or an ultrafiltration concentrator. As the name... prep to the desired concentration using these columns. If you are conjugating your antibody in a downstream...Current Opinion in Colloid & Interface Science, Volume 19, Issue 5, 2014, Pages 438-449, ISSN 1359-0294...

...protrusions and overlaps (arrows) after blue light illumination. Scale bar 25 µm. Images adapted from Mahlandt...GFP-PASS (green). After 30 minutes of blue light illumination, substantial PA accumulates in the plasma membranes...other enzymes. Conclusion We hope this post has illuminated some of the many tools available for controlling...Hehnly, H. (2023). Rab8, Rab11, and Rab35 coordinate lumen and cilia formation during zebrafish left-right ...

...To acquire these measurements, the cytometer illuminates the passing cell with a laser and detects the...colleagues.   Figure 1: The laser pulse illuminates the cell and is scattered forward at a low angle...represents the time and intensity of the cell’s illumination (see Figure 2). H describes the maximum signal...and exclude any events where multiple cells were clumped together. (B) Instead of FSC, one is free to use...

...solution to avoid autofluorescence is offered by bioluminescent proteins. These probes generate light by a ...there is no need to excite the sample. Although bioluminescent probes are usually dim relative to fluorescent...palette of Nano-lanterns for real-time multicolor luminescence imaging." Proceedings of the National Academy...PMC4394297. 16. Iwano, Satoshi, et al. "Single-cell bioluminescence imaging of deep tissue in freely moving animals...

...Dec;11(12):1261-6. doi: 10.1038/nmeth.3147. Bioluminescent pH sensor Understanding intracellular pH regulation...pRSETb-pHlash and pcDNA3.1+-pHlash, that use Bioluminescence Resonance Energy Transfer (BRET) were developed...simultaneously acquired from the the fluorescent and luminescent siganls and a ratio is calculated (the BRET ratio...voltage sensitivity and speed, and an endoplasmic reticulum export motif and trafficking sequence for improved...dynamics and interactions between the endoplasmic reticulum (ER), mitochondria, cytoskeleton, and endosomes...PMID: 26456817. Nano-lanterns for multicolor luminescence imaging  Fluorescent imaging techniques have...The Nano-lanterns rely on the principle of bioluminescence resonance energy transfer (BRET), whereby photons...

...the plasmid DNA from the column using a particular buffer or water. See column manufacturers for more detail...DNA; filter the remaining plasmid DNA through a column that binds the plasmid DNA and lets other materials... detail. Find columnless purification protocols on our website. 6. Verify the plasmid After purifying ...