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Blocking Buffer for Fluorescent Western Blotting

BR, Lbl, WB 1 X
Catalog No. ABIN925618
  • Application
    Blocking Reagent (BR), Labeling (Lbl), Western Blotting (WB)
    Purpose
    Blocking Buffer for Fluorescent Western Blotting
    Specificity
    Blocking buffer was prepared using ultra pure reagents dissolved in pharmaceutical grade water (WFI) and consists of a proprietary protein formulation in TRIS buffered saline at pH 7.6 with thimerosal added as an antimicrobial agent.
    Characteristics
    Western Blot Blocking Solution is specifically designed for western blotting using fluorochrome conjugated antibodies. Pure nitrocellulose membrane is recommended for maximum performance. Other membranes, such as PVDF or nitrocellulose embedded in a support can be used, but may generate elevated backgrounds. Protein should be transferred from gel to membrane using standard protocols.
    Blocking buffer can be used for membrane blocking and to dilute both primary and secondary antibodies.
    Western Blot blocking buffer is suitable for use with fluorescent western blot imaging systems produced by Bio-Rad Laboratories, GE Healthcare, Alpha Innotech, FujiFilm Life Science, Licor Biosciences, UVP and Syngene.
    Purification
    This product was aseptically filtered through a Millipore 0.22 micron filter into clean, pre-sterilized containers. The product was tested on trypticase soy agar for 24 hours, 48 hours and 72 hours and was found to be negative for bacteria.
    Sterility
    Sterile filtered
  • Application Notes
    Western Blot : User Defined
    Other Dilution: Optimum performance is achieved using this product undiluted. However, this blocking solution can often be diluted 1:1 in TBS without a significant loss of performance.
    Comment

    Synonyms: Multiplex Blocking Buffer, Fluorescent Blocking Buffer, Blocking Solution, Blocking Buffer Western Blot, IRDye Western Blot Blocking Buffer, Alexa Dye Blocking Buffer, DyLight Blocking Buffer

    Protocol
    Fluorescence technology is widely used to detect proteins in both the visible and near-infrared ranges. This product allows for superior signal detection and lower background noise when fluorochrome conjugated antibodies are used to visualize proteins in western blotting and other applications. Antibody conjugates prepared with IRDye® 800 and IRDye® 700DX (Licor), Cy2™, Cy3™, Cy3.5™, Cy5™ and Cy5.5™ (GE Healthcare), DyLight™405, DyLight™ 549, DyLight™ 649, DyLight™ 680, and DyLight™ 800 (Thermo Fisher/Pierce) and Alexa Fluor® 488, Alexa Fluor® 532, Alexa Fluor® 546, Alexa Fluor® 647 and Alexa Fluor® 680 (Invitrogen/Molecular Probes) have been validated on various platforms using this product with superior results compared to other commercially available products. In the infrared range, where little to no autofluorescence occurs, specific signal is sharply evident from any background giving the best possible signal-to-noise ratio. This allows for detection levels in the picogram range which rivals the sensitivity of chemiluminescence on film for western blotting. Superior results are also seen when this product is used for simultaneous labeling (multiplex) in western blots or microscopy using various fluorochrome combinations for multicolor imaging. Membranes blocked with this product can be dried and are very stable. Membranes that are stored protected from light can be re-washed and/or rescanned.
    Restrictions
    For Research Use only
  • Format
    Liquid
    Concentration
    1 X
    Preservative
    Thimerosal (Merthiolate)
    Precaution of Use
    This product contains thimerosal (merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Storage
    4 °C
    Storage Comment
    Store Blocking Buffer at 4° C prior to opening. DO NOT FREEZE.
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