Render Target: STATIC
Render Timestamp: 2024-12-13T10:44:56.497Z
Commit: 611277b6de3cd1bb065350b6ef8d63df412b7185
XML generation date: 2024-09-27 20:01:25.357
Product last modified at: 2024-12-11T21:15:08.753Z
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PDP - Template Name: Buffer Kit
PDP - Template ID: *******aace572

Blue Loading Buffer Pack #7722

    Product Information

    Product Usage Information

    1. Prepare fresh 3X Reducing SDS Loading Buffer by adding 1/10 volume 30X DTT Reducing Agent to 1 volume of 3X SDS Loading Buffer.

    2. Dilute 3X SDS Loading Buffer to a 1X solution using ddH2O. This product supplies enough 3X material to make 24ml of 1X solution.     

    3. Aspirate media from cultures; wash cells with 1X PBS; aspirate.

    4. Lyse cells by adding 1X SDS Loading Buffer (100 µl per well of 6-well plate or 500 µl per plate of 10 cm2 plate). Immediately scrape the cells off the plate and transfer the extract to a microcentrifuge tube. Keep on ice.

    5. Sonicate for 10-15 seconds to shear DNA and reduce sample viscosity. 

    6. Heat a 20 µl sample to 95-100ºC for 5 minutes; cool on ice.

    7. Microcentrifuge for 2-5 minutes.

    8. Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).

    Solutions and Reagents

    Quantity 8 ml 3X Blue Loading Buffer and 1 ml 30X Reducing Agent3X Blue Loading Buffer Pack: 187.5 mM Tris-HCl (pH 6.8 at 25ºC), 6% (w/v) SDS, 30% glycerol and 0.03% (w/v) bromophenol blue. (Store at room temperature.)  30X Reducing Agent: 1.25 M dithiothreitol (DTT) (Store at -20ºC.)

    Storage

    Store at Room Temperature and -20°C.
    3X Blue Loading Buffer: 187.5 mM Tris-HCl (pH 6.8 at 25°C), 6% (w/v) SDS, 30% glycerol and 0.03% (w/v) bromophenol blue. (Store at room temperature.)
    30X Reducing Agent: 1.25 M dithiothreitol (DTT) (Store at –20°C.)

    Product Description

    The Blue Loading Buffer Pack is used to lyse cells for Western blotting analysis
    For Research Use Only. Not For Use In Diagnostic Procedures.
    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit our Trademark Information page.