Sperm Antibody
Positive/Negative Serum ControlsFOR RESEARCH USE ONLY
not for use in diagnostic proceduresDescription:
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.
Uses:
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.
Specifications:
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.
Recommendations:
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.
Caution:
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.
Storage:
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
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