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Molecular Probes™ Click-iT™ Plus Alexa Fluor™ 647 Picolyl Azide Toolkit

Catalog No. C10643
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Description

Click-iT™ Plus Alexa Fluor™ 647 Picolyl Azide Toolkit

Click-iT™ Plus Alexa Fluor™ picolyl azide toolkits contain everything you need to perform copper-catalyzed click reactions with compounds that are sensitive to copper, while retaining all of the benefits of the standard azide/alkyne click reaction. Picolyl azides react efficiently with chelated copper, so free copper in the click reaction can be minimized, protecting against undesired copper side reactions with proteins (e.g., GFP, RPE), nucleic acids (e.g., RNA, oligos), and even small molecules (e.g., phalloidin).

  • Flexible—optimize your own Click–iT™ reaction for the detection or labeling of any alkyne-containing biomolecule in vitro or in cells and tissue samples
  • Bright and sensitive—superior fluorescent Alexa Flour™ dyes combined with the efficiency of picolyl azide
  • Convenient—contains all the buffers and reagents needed to create the optimal click reaction

    Each Click-iT™ Plus Picolyl Azide Toolkit includes 100μg of Alexa Fluor™ dye-picolyl azide and all of the reagents required to create a reaction cocktail with the optimal copper and chelate concentration for any click chemistry-based labeling or detection application. Guidelines for designing and optimizing the picolyl azide/alkyne click reaction of your choice are provided in the product manual. Additionally Click-iT™ Plus picolyl azides offer increased sensitivity and faster reaction times compared to standard azides using identical reaction conditions.

    • Specifications

    Description Click-iT™ Plus Alexa Fluor™ 647 Picolyl Azide Toolkit, 5mg, Solid, 650nm Emission, 670nm Excitation, 1 Year Shelf Life, 2 to 8°C Storage Requirements, Less Hazardous Green Features, Used for Labeling Chemistry
    For Use With (Application) Labeling chemistry, cell analysis, click chemistries, glycoprotein analysis with click chemistry, metabolic labeling and nascent protein synthesis with click chemistry, cell proliferation with click chemistry, nascent RNA detection and capture with click
    Kit Contents 1 x 100μg Alexa Fluor™ dye-picolyl azide, 1 x 4mL Click-iT™ reaction buffer, 1 x 1.5mL copper (II) sulfate, 1 x 500μL copper protectant, 1 Click-iT™ buffer additive
    Product Type Click-iT™ Plus Alexa Fluor™ 647 Picolyl Azide Toolkit
    Quantity 1 Kit
    Content And Storage
    • 1 x 100 μg Alexa Fluor™ dye-picolyl azide
    • 1 x 4 mL Click-iT™ reaction buffer
    • 1 x 1.5 mL copper (II) sulfate
    • 1 x 500 μL copper protectant
    • 1 Click-iT™ buffer additive
    Store at 2 to 8°C
    Excitation/Emission 650/670
    Product Line Click-iT
    Label or Dye Alexa Fluor 647
    Format Solid

    Frequently Asked Questions (FAQs)

    What is the fluorescence excitation and emission maxima of Alexa Fluor 647 dye?

    Alexa Fluor 647 has an excitation/emission maxima of 650/670 nm.
    What are the main characteristics of a Click-iT reaction?

    Click reactions have several general characteristics: the reaction is efficient, no extreme temperatures or solvents are required, the reaction is complete within 30 minutes, the components of the reaction are bio-inert, and perhaps most importantly, no side reactions occur-the label and detection tags react selectively and specifically with one another. This final point is a key advantage of this powerful detection technique; it is possible to apply click chemistry-labeled molecules to complex biological samples and detect them with unprecedented sensitivity due to the extremely low background of the reaction.

    I will be performing a cell proliferation assay using Click-iT EdU kit. At what point can I stop overnight, or do I have to perform all the steps continuously?

    One may store the sample after fixation overnight in PBS at 4oC. For longer storage (<1 week) , store in buffer with 1-2% formaldehyde or in formalin to limit microbial growth. If you use sodium azide as a microbial inhibitor, it must be completely removed prior to the Click-iT reaction.
    Can I combine Click-iT or Click-iT Plus reactions with phalloidin conjugates used for actin staining?

    We do not recommend using phalloidin conjugates for staining actin in combination with traditional Click-iT or Click-iT Plus reactions since phalloidin is extremely sensitive to the presence of copper.

    For staining actin in combination with traditional Click-iT or Click-iT Plus reactions, we recommend using anti-α-actin antibodies for staining actin in the cytoskeleton. You can find a list of our actin antibodies here.

    Another option would be to use the Click-iT Plus Alexa Fluor Picolyl Azide Toolkit (Cat. Nos. C10641, C10642, C10643). These Click-iT Plus toolkits provide Copper and Copper protectant separately which makes it easier to titrate the copper concentration to obtain optimal labeling with minimal copper-mediated damage. You may need to optimize the click reaction with the lowest possible concentration of copper and then perform the phalloidin staining.

    For Research Use Only. Not for use in diagnostic procedures.