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We narrowed to 545 results for: SEC

Showing: 1 - 20 of 545 results

  1. Deisseroth INTRSECT Collection

    Type
    Collection
    ...Depositor Collections INTRSECT Plasmid Collection INTRSECT Plasmid Collection INTRSECT Implementation Plasmids...SOM-IRES-Flp). Image from Fenno et al., 2014. INTRSECT INTRSECT (intronic recombinase sites enabling combinatorial...Find intersectional expression plasmids that combine multiplexed recombinases, including Cre, Flp, and...neurons defined by a single gene or projection). Intersectional expression approaches combine multiplexed recombinases... multiple parameters. Figure 1: Examples of intersectional cell population definitions using recombinases...recombinases. A) Cre and Flp are multiplexed to enable intersectional viral targeting of populations expressing only...recognition sequences within synthetic introns. INTRSECT was first shown as a proof-of-concept targeting...

  2. Worm Expression Resources

    Type
    Collection
    ... links to protocols, can be found below: SapTrap-SEC kit for C. elegans genome engineering - Bob Goldstein...incorporate a self-excising drug selection cassette (SEC). More information regarding this kit can be found...

  3. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

    Type
    Blog Post
    ...cassette (SEC). To make it possible fluorescently label proteins in C. elegans, they inserted the SEC into...their Roller phenotype. If SEC is inserted at the 5’ end of a gene, the SEC separates the targeted gene...to remove the SEC and after 5-6 days of growth, wild-type animals that have undergone SEC excision are ...the Genetics Society of America. To use this SEC-based system, first design and clone 500-700 bp homology...homology arms for your gene of interest into the FP-SEC vector using Gibson assembly. This construct will... attempt for 6/7 genes. The data are clear - the SEC-based system is both simple to use and highly efficient...genome modifications, including targeted mutations. SEC-based systems may prove effective in other species...

  4. Antibodies 101: Secondary Antibodies

    Type
    Blog Post
    ...about secondary antibodies. While a primary antibody binds to the protein of interest, a secondary antibody...conjugated to the secondary antibody instead of the primary antibody. Since multiple secondary antibodies can...with BioRender.com.    How do secondary antibodies work?  Secondary antibodies are antibodies generated...While most secondary antibodies recognize broad isotype classes, it is possible to get secondary antibodies... conjugated secondary antibody, which will bind to the primary antibody. Perform a second series of wash.... Primary antibodies are represented in blue; secondary antibodies are represented in purple; signaling...antibody’s Y) is large enough to allow multiple secondary antibodies to bind to a primary antibody without...

  6. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    ...techniques have in common? They all typically require secondary antibodies, frequently of the mouse or rabbit ...Görlich lab and their anti-mouse and -rabbit IgG secondary nanobodies toolbox. Nanobodies are like tiny antibodies...the following to occur: A) Multiple polyclonal secondary IgGs can bind to different epitopes of a primary...antibodies are large and bivalent, primary and secondary antibodies cannot be incubated together without... least from different IgG subclasses, so that secondary antibodies exclusively recognize only one primary...yielding similar signal strength to a standard IgG secondary antibody (Pleiner et al figures 4A-C). And since...targets (Pleiner et al figure 4D). Unlike large secondary antibodies, small nanobodies do not oligomerize...

  8. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...purification using qPCR or digital droplet PCR. Insect cells Insect cells are a less common option for the generation... viruses that specifically infect insects. This is what makes insect cells a popular choice for protein...Mammalian versus insect cells for viral production Consideration Mammalian cells Insect cells Viruses...different cell types, including plants, mammals, and insects. In this post, we will compare two of the most ...most widely used systems, mammalian and insect cells. Whether you’re new to in-house virus production or ...passage (transfection). This differs from typical insect cell preparations, which we talk about below. Processing...protein production. The most commonly used cells in insect viral production are Sf9 cells, which are derived...

  9. Genomic Deletions in Mammalian Cell Lines

    Type
    Collection
    ...parameters: 95 °C for 15 min, 35 cycles of (95 °C for 30 sec, 60 °C for 1 min, 72 °C for 1 min), and 72 °C for...Along with the video, you can find the protocol section from the original publication below. You can also...construct. Electroporate cells with 250 volts for 5 msec in a 2 mm cuvette using an electroporation system...

  10. AAV ddPCR Titration

    Type
    Protocol
    ...Vortex primers, probe and master mix for 15 sec then spin 10 sec in a mini centrifuge and place on ice. Wipe...Vortex the master mix for 15 sec and spin in a mini centrifuge for 10 sec before use. Place an 8-well ...Cycling Step Temperature (°C) Time (min) Ramp Rate (°C/sec) # Cycles Denaturation 95 10 2 1 Denaturation 95 ...cartridge with the DG8 gasket, making sure that it is secure. Transfer the cartridge holder to the droplet generator...

  11. Lentivirus ddPCR Titration

    Type
    Protocol
    ...primers/probe mixes and master mix for 15 sec then spin 10 sec in a mini centrifuge and place on ice. Wipe...Vortex the master mix for 15 sec and spin in a mini centrifuge for 10 sec before use. Place an 8-well ...Cycling Step Temperature (°C) Time (min) Ramp Rate (°C/sec) # Cycles Enzyme Activation 95 10 2 1 Denaturation...cartridge with the DG8 gasket, making sure that it is secure. Transfer the cartridge holder to the droplet generator...

  12. Protocol - Bacterial Transformation

    Type
    Protocol
    ...of the tube into a 42°C water bath for 30-60 secs (45 secs is usually ideal, but this varies depending ... you plate 50 μL on one plate and the rest on a second plate. This gives the best chance of getting single...

  13. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ... Centrifuge in microfuge tube at 10,000 g for 30 sec. Pour off the supernatant, being careful not to disturb...is not available. Vortex microfuge tube for 30-60 sec. Centrifuge the tube for 5 min at room temperature...

  14. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...using SYBR detection: 98 °C 3 min / 98 °C 15 sec / 58 °C 30 sec / read plate/ repeat 39x from step 3 / melt...a single peak should be seen. The presence of a second peak at a temperature of ~70–75 °C usually indicates...

  15. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ... in oxidizing environments when targeted via the Sec translocon." Traffic 12.5 (2011): 543-548. Pubmed...compose the secretory pathway, endocytic vesicles, mitochondria, lysosomes or they are secreted into the ... blue fluorescent protein, secBFP2, for studies in the eukaryotic secretory pathway."Biochemical and biophysical...residues form inappropriate disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally, FPs ...and a sequence intended to localize them to the secretory pathway, a dark pool that is mislocalized to the...misfolded, blocked from proceeding through the secretory pathway, and retained in the ER. FPs have been...variety of cellular compartments including the secretory pathway (Figure 2), the inner membrane space of...

  16. AAV Production in HEK293 Cells

    Type
    Protocol
    ...ratio). Shake the bottle up/down vigorously for 30 sec (it’s okay to make bubbles). Incubate at RT for 15... all resuspended cell pellets and sonicate 5 x 1 sec pulses with at least 5 minutes on ice between each...

  17. Antibody Guide

    Type
    Collection
    ...similar factors. Secondary antibodies Secondary antibodies, often called secondaries, are usually much...and a conjugated secondary antibody specific to your primary antibody. Multiple secondary antibodies will...if your fluorophore is bound to a secondary antibody, the secondary antibodies must be able to distinguish...Therefore, secondary antibodies are species specific and are often referred to by [secondary source] [primary...[primary source secondary recognizes], e.g. , “goat anti-rabbit.” Choose a secondary that recognizes your...applications, and is covered in the following Isotypes section. The variable region of the antibody is the antigen...the amplification of signals by having multiple secondary antibodies binding to a primary antibody which...

  18. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...Pro-Tip It is a good idea to microwave for 30-45 sec, stop and swirl, and then continue towards a boil...the sample is unloaded, push the pipettor to the second stop and carefully raise the pipette straight out...

  20. Immunology Research Plasmids and Resources

    Type
    Collection
    ... the secretion of antibodies, and also function as antigen presenting cells. B cells in secondary lymphoid...chromogranin A (parathyroid secretory protein 1) CGA CHGB chromogranin B (secretogranin 1) SCG1 CKLF chemokine-like... SDS, SWDS SCG2 secretogranin II (chromogranin C) CHGC, SN, SgII SCGB3A1 secretoglobin, family 3A, member...HIN1, LU105, MGC87867, PnSP-2, UGRP2 SCT secretin - SCTR secretin receptor SR SCYE1 small inducible cytokine...SDCN, SYND3 SDC4 syndecan 4 MGC22217, SYND4 SECTM1 secreted and transmembrane 1 K12 SEMA3A sema domain,... a recurrent infection and rapidly initiate a secondary immune response. Browse Plasmids for Immunology...
Showing: 1 - 20 of 545 results