Specificity of the generated antibodies against human ADAM2.

<p><b>A</b> and <b>C</b>. Testicular germ cells were isolated from monkey testis, boiled in 3% SDS and 5% β-mercaptoethanol, subjected to SDS-PAGE, and blotted with <b>A</b>. anti-hADAM2-Cys and <b>C</b>. anti-hADAM2-CyT antibodies. <b>B</b> and <b>D</b>. Human testis was lysed with 0.1% NP-40 buffer, and Western blot analysis was performed using <b>B</b>. anti-hADAM2-Cys and <b>D</b>. anti-hADAM2-CyT. Glutathione S-transferase or GST recombinant ADAM2 (GST-hADAM2-Cys and GST-hADAM2-CyT) proteins were added to the buffer containing the primary antibodies during Western blot analysis. <b>E</b> and <b>G</b>. The supernatants from the monkey testis lysates were boiled in 3% SDS with (lane 1) or without (lanes 2 and 3) 5% β-mercaptoethanol, and blotted with <b>E.</b> anti-hADAM2-Cys and <b>G</b>. anti-hADAM2-CyT. <b>F</b> and <b>H</b> Human testis supernatants were boiled in 3% SDS and with (lane 1) or without (lanes 2 and 3) 5% β-mercaptoethanol, and immunoblotted with <b>F</b>. anti-hADAM2-Cys and <b>H</b>. anti-hADAM2-CyT. Experiments were repeated three times. Reduced protein samples were subjected to SDS-PAGE using 8% resolving gels. Abbreviation: GST, glutathione S-transferase; β-ME, β-mercaptoethanol. Molecular masses are presented on the left.