Invitrogen™ Normal Mouse Serum, eBioscience™

Catalog No.	Quantity
50-154-26	500 μL

Catalog No. 50-154-26 Supplier Invitrogen™ Supplier No. 24554494

Description

Mouse Control

Description: This normal mouse serum can be used to inhibit non-specific binding of mouse monoclonal antibodies during staining for flow cytometric analysis. This product can be used with Product # Product # 14-4777-82, Product # 14-4774-82, and Product # 14-797982. It can also be used in other applications which require normal mouse serum.

Normal Sera are non-immune serum samples from goat, human, mouse, rabbit and other species that contain the usual complement of serum proteins, including the various classes of immunoglobulins that exist in healthy animals of the particular species. Normal serum provides a sufficient quantity of endogenous proteins to saturate and block nonspecific binding sites on fixed samples or assay substrates. When used as an antibody diluent, non-immune serum also supplies a native chemical environment for antibodies to function properly without adding foreign protein components. Normal serum is frequently used for blocking or saturating generalized binding interactions for immunodetection methods, especially those involving tissue samples such as immunohistochemistry (IHC). Normal sera are also useful as controls for testing general and specific antibody purification methods.

Specifications

• Content And Storage: 4°C
• Form: Liquid
• Product Type: Normal Mouse Serum
• Quantity: 500 μL
• Concentration: Conc. Not Determined
• Formulation: Whole serum with no preservative
• For Use With (Application): Flow Cytometry, Control
• Isotype: Ig
• Regulatory Status: RUO

Frequently Asked Questions (FAQs)

The antibody I purchased from you says ''concentration not determined'' on the product label and on the product page. How do I determine the concentration?

Some antibodies are provided in non-purified formats, such as ascites fluid, tissue culture supernatant, or whole serum. These formats contain multiple proteins, which can interfere with accurate protein quantification. Traditional methods like A280 measurement or protein assays such as BCA quantify total protein but do not provide a specific concentration for the antibody in question.

Literature, such as "Antibodies" by Harlow & Lane (1988) (https://antibodiesmanual.org/index.php) (Table 8.2 - Storing and Purifying Antibodies), suggests approximate concentrations of antibodies in these solutions, as shown below. However, please note that these are rough estimates, and actual concentrations can vary based on several factors.

Serum
Total antibody concentration: 10 mg/mL
Specific antibody concentration: 0.1 to 1 mg/mL (a maximum of 10% of the antibody population is expected to be the specific antibody of interest)

Tissue Culture Supernatant (10% FBS)
Total antibody concentration: 1 mg/mL
Specific antibody concentration: 0.05 mg/mL

Tissue Culture Supernatant (serum-free)
Total antibody concentration: 0.01 to 0.05 mg/mL
Specific antibody concentration: 0.01 to 0.05 mg/mL

Ascites
Total antibody concentration: 1 to 10 mg/mL
Specific antibody concentration: 0.9 to 9 mg/mL

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.

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.

Safety and Handling

WARNING: Cancer and Reproductive Harm - www.P65Warnings.ca.gov