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We narrowed to 472 results for: FLU;

Showing: 61 - 80 of 472 results

  1. Immunocytochemistry

    Type
    Protocol
    ...primary antibody against a target protein and a fluorescent secondary antibody....primary antibody against a target protein and a fluorescent secondary antibody. This protocol outlines the... tips and pipettes Rocking platform Tweezers Fluorescent microscope 0.5–10 µL single channel pipette 2...500 µL PBS on a rocking platform. Dilute the fluorescently-labeled secondary antibody to the desired concentration...generally ranges from 1-10 µg/mL. Add 500 µL fluorescently-labeled secondary antibody to the wells and ...cell labeling on a microscope with appropriate fluorescent filters. Tips and Troubleshooting The optimal...

  2. AAV Production in HEK293 Cells

    Type
    Protocol
    ...maintenance phase and do not allow cells to reach 100% confluence (80–90% is ideal). Pass and plate the cells the...from 2 x T-175 flasks. Cells should be at ~80% confluence. For each T-175 flask: Aspirate culture media...resuspend cells from the CS2. Cells should be at ~80% confluence. Aspirate culture media and rinse once with 60...Sample Data Figure 1: HEK293T cells at various confluencies. This AAV production protocol should be started...started with cells that are ~80% confluent (center panel). These cells were imaged with a 10X objective ...

  3. General Transfection

    Type
    Protocol
    ...with fresh media Day 3 or more (am): Observe fluorescence, harvest cells, or perform your experiment Equipment...batch of PEI is prepared, transfect cells with a fluorescent plasmid using a variety of ratios. Check the ...efficiencies as can be seen here by the amount of green fluorescent protein expression (green in the right panels...

  4. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...the individual wells become confluent, expand into larger vessels. A confluent well of a 6-well dish can...can be expanded into a 10 cm dish. A confluent 10 cm dish can be expanded into two 75 cm 2 flasks, etc....

  5. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...the next day they will be approximately 50–60% confluent. For 293T cells this is about 2 × 10 6 cells in...Do not use the medium if the cells are overly confluent or the medium is acidified (i.e., the medium appears...cells have expanded but before they become over-confluent, trypsinize the cells and expand them to larger...

  6. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...cells from incubator. The cells should be 50-80% confluent and in DMEM that does not contain antibiotics....The target cells should be approximately 80-90% confluent. c. Dilute puromycin in the preferred culture ...: b. Target cells should be approximately 70% confluent. Change to fresh culture media containing 8 μg...

  9. Lentivirus Production

    Type
    Protocol
    ...batch of PEI is prepared, transfect cells with a fluorescent plasmid using a variety of ratios. Check the ...

  10. CRISPR Library Amplification

    Type
    Protocol
    ...different concentrations of DNA. In our hands fluorescent dye methods tend to report lower values than ...

  11. Lentivirus ddPCR Titration

    Type
    Protocol
    ... Colony Formation Titration Assay Lentivirus Fluorescence Titration Assay AAV Titration by qPCR Addgene...

  12. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ...  APCAlexa Fluor 647   Alexa Fluor 700APC-H7APC-Cy7   Many factors can influence the level of... you'll need to pick a fluorophore for each target and ensure the fluorophores, together, are able to ...reporter's fluorescence. Another way to label proteins is through treating cells with fluorophore-conjugated...dead cells. The combination of antibody-fluorophores, fluorescent reporters, and/or dye stains used in your...outlining a few commonly used fluorophores and their emission spectra. Some fluorophores carry their emission ...and RB545. Fluorophores can also come in combination of two conjugated single fluorophores, like APC-Cy7...respective colors of a few commonly used fluorophores. Each fluorophore has a defined emission maximum. Note...

  13. Bacterial Expression Systems

    Type
    Collection
    ... Right Connection Photoactivatable Fluorescent Proteins Fluorescent Protein Timers Return to top Protein...interactions using fluorescence microscopy. FRET relies on the principle that when a donor fluorophore is excited...will be able to detect fluorescence both from the donor and the acceptor fluorophore by only exciting the...ratio Fluorescence (GFP) Escherichia coli Gary Yellen 187836 pVoPo-01 Promoter activity Fluorescence (mCherry...activity Fluorescence (mCherry) Escherichia coli Mattheos Koffas 173481 HC-M ATP Fluorescence (GFPmut2...activity Fluorescence (mRFP1) Escherichia coli Mary Dunlop 124605 pSCM001 Cytoplasmic pH Fluorescence (mCherry...Promoter activity Fluorescence (turbo-365) Mycobacterium sp. Tanya Parish 26156 pMV306hsp+FFluc Promoter activity...

  14. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...fusion protein called CalFlux VNT. CalFlux VNT is composed of Venus fluorescent protein, the Troponin ...NLuc to fluorescent proteins to make better reporters and the second couples NLuc to fluorescent proteins...better biosensors. NanoLuc®-fluorescent protein fusions for imaging Fluorescent proteins are fantastic imaging...couple Nluc with a red-shifted fluor. Here, NLuc internally excites the fluor through a process known as ...BRET). The fluor then emits light at a tissue-penetrating wavelength above 600 nm. LumiFluors Schaub, F.X...more penetrative orange fluorescent protein fusion (OgNLuc). Both fluorescent proteins can be excited ... the lab has dubbed these fusions LumiFluor Reporters. The LumiFluor reporter constructs were designed...

  15. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    ...the associated fluorophore or stained with the associated fluorochrome, emits a fluorescent signal that ...the level of fluorescence. A cell expressing high levels of the fluorophore or fluorochrome-stained protein...antibody conjugated fluorophores Fluorescent antibodies get around the challenges of fluorescent protein fusions...such as fluorescent microscopy must be used. To learn more about the most suitable fluorescent microscopy... have a variety of fluorescent light sources. When interrogated with a fluorescent light source, a cell... to a fluorochrome. In indirect staining a primary antibody binds to a target and a fluorochrome-conjugated... panel of “fluorescence-minus one” controls, samples stained with all of the fluorochromes in the experiment...

  16. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ...non-radiatively from an excited molecular fluorophore (the donor) to another fluorophore (the acceptor) by means of ...the two fluorophores used are in close vicinity. Thus, the distance between the two fluorophores and their... of a fluorophore pair, donor:acceptor stoichiometry, and cross-talk between the two fluorophore colours... filters chosen for the donor fluorescence and the acceptor fluorescence. Additionally, this method could... the fluorescence decay time of the donor. When FRET occurs between the pairs, donor fluorescence is quenched...factors, such as pH or autofluorescence background, can change the fluorescence decay time and have to...course talking about the now well-known FRET, aka Fluorescence (Förster) Resonance Energy Transfer, technique...

  17. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    ...for autofluorescence extraction Cells produce a low level of fluorescence called autofluorescence (you...to identify a fluorophore, spectral flow cytometers use all detectors for all fluorophores. This allows...tool called “autofluorescence extraction” which allows the user to use the autofluorescence in a sample...sample as a fluorescent marker. Given that the level of autofluorescence across various tissue types varies...you wish to use the autofluorescence extraction feature. Although autofluorescence extraction is an optional...samples stained with a single fluorophore. They’re required for every fluorophore you use in your panel, whether... the spectral signature of a fluorophore itself from the autofluorescence of the beads or cells. Overall...

  18. Antibody Guide

    Type
    Collection
    ... curve with known amounts of protein. Fluorophores Fluorophores are light-sensitive molecules which emit...cell or tissue sample. Figure 6: Immunofluorescence, in which fluorophore-conjugated primary antibodies ...label cells in a sample. Immunofluorescence (IF) This method uses fluorophore-conjugated antibodies to...IF uses fluorophore color to differentiate between the targets of interest, if your fluorophore is bound...wells) by fluorescence activated cell sorting (FACS). Figure 7: Flow cytometry uses fluorescently-labeled...the fluorophores used is important. It is best to inversely pair the brightness of the fluorophore with...If a bright fluorophore is on an abundant target, it obscures fainter signals. Fluorophores can emit signals...

  19. Brain Initiative Collection

    Type
    Collection
    ...genome encoding Cre-dependent expression of the fluorescent protein EYFP from the CAG promoter Viviana Gradinaru...An AAV genome that ubiquitously expresses the fluorescent protein eYFP from the CAG promoter Viviana Gradinaru...Cation channelrhodopsin ChrimsonR fused to mRuby2 fluorophore and targeted to the neuronal soma and proximal... AAV production plasmid encoding for Archon1 fluorescent voltage reporter, Cre-dependent expression Edward...

  20. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ... blog Fluorescent Proteins 101: Fluorescent Biosensors Resources on Addgene.org Fluorescent protein...area with lots of bright new plasmids is our Fluorescent Biosensors Collection — scientists are always...measure! Let’s take a look at some of the hottest fluorescent biosensors new to Addgene this year. Feel the...Hiroshi Hibino and colleagues devised a “Modular fluorescence unit fused with TRansmembrane region-to-IntrAcellular...region-to-IntrAcellular loop linkers” (MTRIA for short), where a fluorescent protein is inserted into one of the loop regions...  Find MTRIA sensors here!   Ino D, et al. A fluorescent sensor for real-time measurement of extracellular...co-expressing a cytosolic ZtSH2 reporter, they observe fluorescence localizes to the plasma membrane only when the...
Showing: 61 - 80 of 472 results