We narrowed to 295 results for: SAM activators
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TypeBlog PostPublishedFeb. 4, 2025, 2:15 p.m....wide variety of screens, including interference, activation, and knockout. Most CRISPR screens rely on ...expression of lentiviral vectors make them an attractive option. However, safety requirements and expression...vivo. The general process of these screens is the same: Deliver CRISPR library to cells of interest. Select...Low, S. H., François, A., Suárez-Amarán, L., & Samulski, R. J. (2022). AAV vectors: The Rubik’s cube of...
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AAVs in Retinal Gene Therapy
TypeBlog PostPublishedJuly 31, 2018, 12:55 p.m....back, better and safer. It is now one of the most active and exciting areas of science. This is hopefully...retinal cells - AAV - were becoming available. At the same time advances in human genetics led to the identification...24664759. PubMed Central PMCID: PMC4267879. 10. Samulski RJ, Muzyczka N., 2014. AAV-Mediated Gene Therapy... -
Plasmids 101: Secondary Nanobody Toolbox
TypeBlog PostPublishedFeb. 27, 2018, 2:04 p.m....State University Use Nanobodies to Control Protein Activity Learn more about protein tags Read about E. coli...figures 4A-C). And since they have limited cross-reactivity between different IgG subclasses and different...primary antibody-nanobody mix can be added to your sample, thus eliminating the secondary antibody incubation...568, or Alexa Fluor 647 before application to the sample. Staining with this multi-color staining workflow... -
Tips for Using FRET in Your Experiments
TypeBlog PostPublishedNov. 5, 2014, 3:41 p.m....used to measure conformational changes, cleavage activity using FRET-based biosensors, and interactions ...What is FRET? In their JCB 2003 paper, Sekar and Periasamy defined FRET as "a distance-dependent physical...341-50. doi: 10.1002/bies.201100098. Sekar RB, Periasamy A. Fluorescence resonance energy transfer (FRET... -
Antibodies 101: Reading a Flow Plot
TypeBlog PostPublishedFeb. 6, 2024, 3:13 p.m.... dendritic cell function and increased B cell activation. PLOS ONE, 12, e0180664. https://doi.org/10.1371... are represented as a series of dot plots in the same order. However, in an actual flow cytometry run,...parallel. Let’s imagine we’re looking for cells in a sample that are negative for marker A and positive for... proxy for nucleic complexity of the cell. The sample will have been labeled using a panel of markers... dots. In a contour plot, each line contains the same number of events. Thus, contour lines that are close... -
Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View
TypeBlog PostPublishedOct. 19, 2017, 1:54 p.m....the nucleus where they exert their recombinase activity (Logie and Stewart 1995; Metzger et al. 1995)....reintroduction, inversion and re-inversion happen at the same probability. Continuous inversion of the DNA sequence...recombination can only occur between target sites of the same sequence in FLEx vectors, the DNA sequence that ...manipulations in vivo, FLEx vectors expressing the light activatable ion channel under the control of Cre recombinase...neuron and its synapse should be visualized at the same time from two different plasmids this approach becomes...connectivity before and after the gene knockout in the same mouse. Scientists always strive for more.... -
Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection
TypeBlog PostPublishedDec. 12, 2013, 3:35 p.m....releasing simply plug into this existing economic activity. People are using BioBrick parts to make more ...BioBricks Foundation to distribute the physical DNA samples, for when that's the preferred mode of receiving... -
pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs
TypeBlog PostPublishedDec. 14, 2017, 2:08 p.m....human peripheral blood using a combination of activated T cells and Sendai virus. Nat Protoc, 2012. 7(...blood sample, in the best-case scenario [7]. In the clinical setting where patient blood samples are limited...grade hiPSCs from easily accessible patient blood samples. All pCXLE vectors listed below in Table 1 are ... -
Evolution of Lab Techniques
TypeBlog PostPublishedJune 21, 2016, 2:30 p.m....synthesized by electronically recording fluorescence activity as fluorescently labeled nucleotides are incorporated...membrane for analysis through hybridization of a radioactively or fluorescently labeled probe. If the DNA is...searching for), then the probe will bind to the sample thereby labeling it. The label can then be visualized...visualized by autoradiography and X-ray film for radioactively labeled probes or by the presence of fluorescence...strength of the signal in comparison to a control sample. Southern blotting applications range from confirming... MstII and southern blotting are conducted on a sample from an individual without sickle cell anemia, ... largely replaced by real-time PCR to answer the same experimental questions. Real-time PCR detects and... -
Antibodies 101: Introduction to Gating in Flow Cytometry
TypeBlog PostPublishedApril 4, 2024, 1:15 p.m....gate to focus your analysis on what’s biologically active. After that, the next gates fully depend on your...When using flow cytometry to analyze your samples, it is necessary to set up a sequence of gates to be...the area under the resulting curve. By default, a sample on the flow cytometer (before you set the first...Practical example: lymphocytes in a mouse tumor sample You made it! So much about the theoretical background...gated out my lymphocyte population from a tumor sample. Many cell types can be found in the tumor microenvironment...proportion between H and A (or W and A) within the same type of scatter. Here I employed an FSC-H / FSC-...pattern of positive vs. negative signal remains the same (Figure 5). A. ... -
Hot Plasmids Spring 2024
TypeBlog PostPublishedMay 23, 2024, 1:15 p.m....for a range of applications, including CRISPR activation by dCas9-VPR, prime editing, and delivery of ...#211756 (left panel) or #211757 (middle panel). Samples were also immunoblotted against vinculin as a loading...during action potentials of fast-spiking neurons. The same A-to-D mutation in the related voltage indicator... -
Getting the Most from Your Lentiviral Transduction
TypeBlog PostPublishedApril 7, 2016, 2:30 p.m.... nucleic acid or express transgenes as well as actively dividing cells resulting in lower lentiviral titers.... Know your target cells Labs tend to follow the same general procedure for lentiviral infections, produce...but also between different lots of serum from the same source. Labs that routinely produce lentivirus may... -
Fluorescent Proteins 101: Fluorescent Protein Timers
TypeBlog PostPublishedMay 4, 2017, 2:30 p.m....changes in oxygen, temperature, and pH, but timer activity can be calibrated to these different conditions...and look at them under a microscope, fractionate samples to isolate particular organelles and their contents... -
Degrading DNA with Cascade-Cas3
TypeBlog PostPublishedFeb. 11, 2025, 2:15 p.m....non-target strand. The helicase domain is then activated, and Cas3 begins unraveling the double-stranded...using Cas9, two or more gRNAs must be used along the same chromosome. During DNA repair, there is a chance... -
Plasmids 101: Protein tags
TypeBlog PostPublishedDec. 11, 2014, 4:26 p.m....various mutations used to increase its stability activity, can be readily purified using plasmids found ...protein since the enzyme cleaves the SUMO tag in the same manner as it would have in the context of a cell... -
Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More
TypeBlog PostPublishedDec. 2, 2014, 7:23 p.m.... However, at the same time, it's fun to be involved in these professional activities because I can contribute... -
Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor
TypeBlog PostPublishedJuly 23, 2019, 12:46 p.m....It was discovered as an antigen upregulated on activated lymphocytes and is commonly used to enrich adult...Deverman group then looked to see if PHP.B required the same receptors that its parental AAV9 relied on for transducing... -
Hot Plasmids: Summer 2024
TypeBlog PostPublishedAug. 27, 2024, 1:15 p.m....dCas9, zinc finger proteins, and transcription activator-like effectors (TALEs), with fifteen total variants...protect fragile samples during cryo-EM grid plunge freezing By Rachel Leeson Cryo-EM samples can be damaged...and structure determination LEA proteins protect samples during cryo-EM grid freezing Engineered and evolved...antibodies for marking astrocytes MagIC-Cryo-EM for sample enrichment and structure determination By Mike ...enrichment streamlines the purification and minimizes sample loss while also supporting imaging, making it easy...cryo-EM grid. Figure 1: MagIC-cryo-EM for sample enrichment and structure determination. A) Schematic...beads for scarce macromolecules in heterogeneous samples. bioRxiv 2024.01.21.576499; doi: https://doi.org... -
An Integrin Antibody Toolkit from IPI
TypeBlog PostPublishedMarch 28, 2024, 1:15 p.m....the integrin from its closed to its open state, activating the receptor. Ligands can bind to integrins in...Cells often express multiple integrins that bind the same ligand, making it difficult to dissect the function...-open. In the two closed states, integrins are inactive, with low affinity for ligands. When integrins...extracellular matrix. During immune responses, this same conformation encourages immune cell adhesion and...IPI antibodies Antibody Target Proteins Reactivity Isotype Source Species Anti-Integrin alpha... -
The PAM Requirement and Expanding CRISPR Beyond SpCas9
TypeBlog PostPublishedAug. 20, 2020, 2:30 p.m.... as increased specificity and lower off-target activity (Hu et al., 2018). In 2020, the Kleinstiver lab...and crRNA (or synthetic gRNA) are derived from the same species. Expanding the CRISPR toolbox The isolation...