Positive/Negative Serum Controls
FOR RESEARCH USE ONLY
not for use in diagnostic procedures
Sperm Antibody Positive Serum: 1 ml heat-inactivated human serum with 0.1% sodium azide. Ready to use.
Sperm Antibody Negative Serum: 1 ml heat-inactivated human serum with 0.1% sodium azide. Ready to use.
Note:These serum controls may look cloudy due to lipemia. Lipemic serum will not interfere with test results.
These products are suitable for use as controls in sperm antibody assays such as ImmunoSpheres®, IBT™, MarScreen®, and others. They cannot be used in an indirect MarScreen® IgA, an indirect MarScreen® IgM, or with an indirect ImmunoSpheres® Anti-IgA or Anti-IgM or an indirect IBT™ Anti-IgA or Anti-IgM.
Direct sperm antibody assays (on sperm): Because of the impracticality of using specimens of antibody-positive sperm as a positive control in direct sperm antibody tests, use serum controls instead. To prepare antibody-positive sperm, start with sperm known to be negative for antibodies. Wash sperm free of seminal plasma. Then incubate the washed sperm with Sperm Antibody Positive Serum at a final dilution of 1:10, as in the protocol for the indirect ImmunoSpheres® or IBT™. Wash the sperm free of Sperm Antibody Positive Serum and then use the prepared sperm as a positive control for antibody-positive sperm.
As an additional control, always prepare antibody-negative sperm at the same time using the Sperm Antibody Negative Serum. To do this, incubate the washed sperm with Sperm Antibody Negative Serum at a final dilution of 1:10. Wash the sperm free of Sperm Antibody Negative Serum and then use the prepared sperm as a negative control for antibody-negative sperm exposed to serum.
Indirect sperm antibody assays (in serum): Follow the manufacturer’s directions for testing serum.
A 1/10 final dilution of the Sperm Antibody Positive Serum will give a greater than 90% total binding of Anti-IgG beads in the indirect IBT™ when incubated for one hour at 37°C using donor sperm. A 1/100 dilution gives a negative result.
A 1/10 final dilution of the Sperm Antibody Negative Serum has no detectable immunoglobulins that will bind to sperm.
The researcher should determine the optimal working dilution(s) of these products. The following information is provided as a guideline to the researcher:
MarScreen®: Dilute serum 1:16 before using in the Indirect MarScreen® IgG. For example, add 20 ul serum to 300 ul sperm washing medium.
ImmunoSpheres® or IBT™: Use serum at a final dilution of 1:10 in either Indirect Anti-IgG test. For example, add 50 ul serum to 400 ul sperm washing medium and 50 ul washed sperm (~50 million motile sperm/ml) suspension.
The Sperm Antibody Positive Serum and the Sperm Antibody Negative Serum have been tested and found negative for HIV antibody and hepatitis B surface antigen, but negative results do not guarantee absence of virus.
Store the reagents at 2 - 8°C. They can be used until the expiration date on each label.
Do not freeze.
Sperm Antibody Serum Controls