Fish Serum Blocking Buffer
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Thermo Scientific™

Fish Serum Blocking Buffer

Thermo Scientific Fish Serum Blocking Buffer is steelhead salmon serum in PBS that is especially useful as a blocking agent in immunohistochemistry (IHC) and other detection methods involving mammalian samples.
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Catalog NumberDescription
37527Fish Serum Blocking Buffer, 500 mL
37527X3Fish Serum Blocking Buffer, 3 x 500 mL
Catalog number 37527
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254.00
Each
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Description:
Fish Serum Blocking Buffer, 500 mL
Request bulk or custom format
Price (USD)
254.00
Each
Add to cart
Thermo Scientific Fish Serum Blocking Buffer is steelhead salmon serum in PBS that is especially useful as a blocking agent in immunohistochemistry (IHC) and other detection methods involving mammalian samples. Fish Serum Blocking Buffer is effective at stabilizing solutions and samples used for antibody-binding interactions, while at the same time minimizing the possibility of cross-reaction with mammalian sample components.
Features of Fish Serum Blocking Buffer include:
• Non-mammalian—fish proteins are less likely to have specific binding interactions with antibodies and other mammalian proteins present in typical methods
• Convenient—filtered and stabilized in PBS for compatibility with most assay systems
• Easy to use—can be used as supplied or diluted up to 10-fold as needed
• Flexible - may be used for many different applications, including as a diluent for antibodies

Fish serum offers several advantages over typical blocking buffers. Because salmon is phylogenetically distant from mammals, which are the source of the antibodies and samples used in most experiments, its serum proteins are less likely to have specific binding interactions with proteins used in the experiments. Fish Serum Blocking Buffer is effective at stabilizing solutions and samples used for antibody-binding interactions while at the same time minimizing the possibility of cross-reaction with mammalian sample components. Try it when use of more-standard sera results in background staining in IHC. In addition, this blocker is effective in various cellular imaging protocols based on visible and near-infrared fluorescence detection. It has been used to decrease background and increase signal-to-noise ratios with near-IR fluorescent probes.

For Research Use Only. Not for use in diagnostic procedures.
  1. Alegria-Schaffer, A., et al. (2009). Performing and optimizing Western blots with an emphasis on chemiluminescent detection. Methods Enzymol. 463:573-99.
  2. Hypolite, J.A., et al. (2001). Am. J. Physiol. Cell. Physiol. 280:C254-64.
  3. Wang, L, et al. (2002). J. Clin. Invest. 110:1175-1184.
Specifications
DescriptionFish Serum Blocking Buffer, 500 mL
Product TypeFish Serum Blocking Buffer
Quantity500 mL
FormLiquid
For Use With (Application)ELISA, Western Blot, Immunohistochemistry (IHC)
Concentration1X
BufferPBS
Blocking AgentFish Serum
Unit SizeEach
Contents & Storage
Upon receipt store at 4°C.

Frequently asked questions (FAQs)

How can I reduce background bands in my Western blot?

Optimize the concentration of primary and secondary antibodies. In some cases, increasing the concentration of blocking agent (BSA or non-fat dry milk) or usiing an alternative blocking solution such as Starting Block or SuperBlock may reduce background signal. After incubation with the primary antibody, wash at least 2 times with TBST (include 0.5 M NaCl in one or more of the wash steps). Avoid Nonidet P40 or Triton X-100 in buffers because protein detection is decreased when these detergents are used.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

What is the concentration of protein in the Fish Serum Blocking Buffer?

This information is proprietary. The protein concentration in the Fish Serum Blocking Buffer has been optimized for direct usage without further dilution, however if desired, the blocking buffer may be diluted with phosphate-buffered saline. For best results, our recommendation is to empirically determine the optimal concentration to use.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

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