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Invitrogen™ BlockAid™ Blocking Solution

Catalog No. B10710
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50 mL
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B10710 50 mL
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Catalog No. B10710 Supplier Invitrogen™ Supplier No. B10710
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Description

BlockAid™ Blocking Solution

BlockAid™ Blocking Solution is superior to commonly used blocking reagents such as bovine serum albumin, normal serum, or casein for use with cells or tissue sections. It is an optimized mix of protein-blocking components, with no dilution required. BlockAid™ Blocking Solution can be used as the initial blocking reagent before primary antibody is applied to minimize non-specific antibody binding, and as a diluent for both the primary and secondary antibody. This reagent is appropriate for cell or tissue immunofluorescence applications. It has also been validated for use with streptavidin conjugates and microsphere-protein conjugates.

Important Features of BlockAid™ Blocking Solution:

  • Excellent background reduction—superior to conventional blocking solutions

  • Ready to use—no dilution or stock preparation required

  • Versatile—use with any primary or secondary antibody, streptavidin conjugate, microsphere protein conjugates, or Qdot™ conjugates

    Minimizing background from non-specific protein binding of antibodies is essential for improved sensitivity and maximum signal-to-background ratios. This is especially crucial when looking for low-expressing antigens, when using samples with high autofluorescence, or for techniques where signals are inherently dim (such as use of directly-labeled primary antibodies or use of super-resolution imaging).

    • Specifications

    Type Blocking Solution
    Product Line BlockAid
    Reagent Type Blocking⁄Background Suppression Reagent
    Content And Storage Store in freezer -5°C to -30°C.
    Quantity 50 mL
    Shipping Condition Wet Ice

    Frequently Asked Questions (FAQs)

    Can BlockAid blocking solution be used as a protein blocking solution for antibody labeling on cell or tissue samples, or does it only work with microspheres?

    BlockAid blocking solution (Cat. No. B10710) is a mix of protein blockers which, when used undiluted or slightly diluted, is as good as or better than any other protein blockers we've tested (such as BSA/normal goat serum). It was developed for use with microspheres, but it is great for cell and tissue blocking. Use undiluted for the initial block, then dilute your primaries or secondaries into it for antibody labeling.
    I used a neuron-specific antibody to label my neurons. How can I reduce non-specific antibody binding?

    A blocking step should be performed to reduce fluorescence due to non-specific antibody binding. A common blocking step is the addition of a 2-5% solution of bovine serum albumin (fraction V defatted BSA). Another approach employs the addition of a 5-10% solution of serum from the species in which the secondary antibodies were raised. For example, when using goat anti-mouse IgG secondary antibodies, samples may be effectively blocked with 5-10% normal goat serum. To further reduce background fluorescence, the Image-iT FX Signal Enhancer can be included as a pre-blocking step to decrease non-specific labeling due to charge interactions between the dyes on the conjugates and the cellular constituents.
    If you are using a secondary antibody make sure that the species of the antibody is not the same as the species of the sample. For example do not use an anti-mouse secondary antibody on mouse tissue.
    Titrate the antibody to the lowest concentration you can use and still get adequate signal.
    Try using a fluorescently tagged primary antibody because it should give reduced background but be aware this can reduce signal intensity.
    What should I use to block my cells for flow cytometry analysis?

    Use serum from the same species as the host species of the secondary antibody for blocking. If the serum is not available, use from 2 to 5% BSA (Fraction V, defatted). If using only a primary antibody, such as directly-labeled mouse primary antibodies, a good blocking reagent is Fc block. CD16 + CD32 Antibody (FRC-4G8) (Cat. No. MFCR004 or MA5-16680) is a low-affinity receptor for the Fc region of immunoglobulin gamma complexes.
    After labeling with my antibody, I am seeing non-specific background binding in my cells or tissue. What could be the cause?

    There can be many causes, including insufficient blocking, too high a concentration of the primary or secondary antibody, or degraded primary or secondary antibody. A “no-primary antibody” control can help determine if the secondary antibody is at fault. Otherwise, we recommend trying more stringent blocking or lower concentrations of primary and secondary antibodies.
    What concentration of my antibody should I use for cell analysis?

    An optimal concentration may be between 1-10 µg/mL for cell and tissue labeling for microscopy, or 0.2-5 µg/mL for flow cytometry. A range of concentrations should be tested to determine what is optimal.
    I know I need to block my samples against non-specific protein binding of my antibodies, but which blocker should I use?

    For blocking cells or tissues you may use 2-5% bovine serum albumin (fraction V, defatted BSA) or 5-10% normal serum of the species matching the host species of the secondary antibody. Other options include a mixture of BSA and serum or other purified proteins. We offer a ready-made blocking solution, BlockAid Blocking Solution (Cat. No. B10710), which is not species-specific. For cell or tissue samples, avoid the use of non-fat dry milk as a blocking agent as it contains a high level of phosphoproteins, histones, and biotin.

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