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Showing: 221 - 240 of 284 results
  1. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    Published
    April 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.                                                                                                        ...
  2. Hot Plasmids Spring 2024

    Type
    Blog Post
    Published
    May 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...
  3. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    Published
    April 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...
  4. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 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...
  5. Plasmids 101: Protein tags

    Type
    Blog Post
    Published
    Dec. 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...
  6. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 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...
  7. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    Published
    March 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...
  8. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    Published
    April 5, 2018, 12:42 p.m.
    ... design has been shown to increase T7 promoter activity. To avoid re-cutting by Cas9 after editing, make...They have successfully used guides targeting the same or complementary strands, as well as guides facing...
  9. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    Published
    Aug. 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...
  10. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    Published
    Jan. 26, 2023, 2:15 p.m.
    ...homology-directed genome editing by catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA. Richardson...by intracellular factors such as nucleases and reactive oxygen species, or external forces such as ionizing... occur; however, if both HJs are resolved in the same manner, this results in a non-crossover event.  ...
  11. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    Published
    Jan. 19, 2021, 2:15 p.m.
    ...effect within the body, or they can be used to activate cellular pathways by binding to cell surface receptors... exact amount of protein in your sample, you’ll compare your samples to a standard curve. This curve is...the known samples is used to calculate the concentration of protein in the experimental samples. Get more...multiple proteins on the same cell simultaneously as well as compare between samples. Learn more about flow...of different isotypes can be used together in the same experiment because the reagents used to detect the... population of antibodies that all recognize the same epitope of the target protein. Most commonly these...expression in cell lysates, whole cells, or tissue samples. Many experiments that use antibodies use a primary...
  12. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    Published
    Nov. 2, 2021, 1:15 p.m.
    ...expressing TH protein indicating that there is promoter activity. Additionally, some neurons express low levels... with the same titer and serotype previously used in a different brain region that the same result will...sites will be flanked by two lox sites now in the same orientation. Next, the loxp site is excised since...since it is between two lox2272 sites in the same orientation preventing the transgene from reverting. Figure...scrambled one pair of lox sites and either used the same sequence to flank a transgene (homologous pairs)...the DIO/FLEx vector. This step is essentially the same as what was used to optimize the serotype, except...
  13. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    Published
    June 1, 2023, 1:15 p.m.
    ...chemogenetic tools, where modification of cellular activity outside of your experiment site or circuit can... different phenotypes of the same cell type, are intermingled in the same brain nucleus: a serotype that...tropism between animal species or even strains of the same species. What works well in a mouse may not work...doi.org/10.3390/cells12050785 Murlidharan, G., Samulski, R. J., & Asokan, A. (2014). Biology of adeno-...Response against the Tetracycline-Dependent Transactivator Correlates with Long-Term Doxycycline-Regulated...10.3390/pharmaceutics13050750 Murlidharan, G., Samulski, R. J., & Asokan, A. (2014). Biology of adeno-...
  14. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    Published
    Sept. 1, 2020, 1:15 p.m.
    ... hybrid vector, as measured by β-galactosidase activity, was 81% of that from a single vector, but significantly...nbt0702-697 Hirsch ML, Agbandje-McKenna M, Jude Samulski R (2010) Little Vector, Big Gene Transduction:...Bellon I, Yin C, Chavala S, Pryadkina M, Richard I, Samulski RJ (2013) Oversized AAV Transductifon Is Mediated...
  15. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ...RNA cleavage, and translational repression or activation: Kellermann, Stefanie J., and Andrea Rentmeister...plasmids used to generate these fusions all use the same restriction enzymes for cloning. Furthermore, some...
  16. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    Published
    Oct. 5, 2021, 1:15 p.m.
    ...far-red genetically encoded indicators of neuronal activity. J Neurosci Methods 362, 109314 (2021). Yu, D...., and deep penetration of red-shifted light. The same NIR FPs and biosensors can be imaged at spatial ...
  17. Building Global Connections with the International Mentorship Program USA-EUROPE

    Type
    Blog Post
    Published
    Nov. 6, 2015, 3:30 p.m.
    ... metastasis as well as teaching and mentoring activities. Follow her on twitter @zcastan or on linkedln... my mentor, since she does research at UK in the same field that I do: prostate cancer. She has guided... Pilot Program 2015-2015 Scholars need strong proactive mentors who have relevant expertise and skills...
  18. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...Griesbeck, Heim, & Tsien, 1999), monitoring kinase activity (Ni, Titov, & Zhang, 2006), quantifying antibody... However, the acceptor doesn’t need to go to the same exact location as the donor. As long as some of ...targeted nearby (such as a neighboring house on the same street as the donor), then FRET measurements are...Ebright, 2001). These Cy/NTA conjugates work the same way as NTA-sepharose beads used for protein purification... binding sites are adjacent to each other on the same protein and a negative control experiment conducted...
Showing: 221 - 240 of 284 results