Human Anti-SAGA-1 Recombinant Antibody (clone RASA) (CAT#: HPAB-0296CQ)

Figure 1 Western analysis of human sperm extracts demonstrating reactivity of RASA with SAGA-1

Chloroform/methanol-extracted spermatozoa were subjected to preparative SDS-PAGE and immunoblot strips were incubated with the S19 mAb (lane 1, 1:1000) or RASA (lane 2, 1:100) followed by goat anti-mouse (1:5000) or anti-E Tag (1:1000) secondary antibodies, respectively. Both mAbs detected the identical polymorphic pattern (~15–25kDa) characteristic of the SAGA-1 glycoprotein. No reactivity was seen with the anti-E Tag antibody alone (lane 3).

Norton, E. J., Diekman, A. B., Westbrook, V. A., Flickinger, C. J., & Herr, J. C. (2001). RASA, a recombinant single-chain variable fragment (scFv) antibody directed against the human sperm surface: implications for novel contraceptives. Human Reproduction, 16(9), 1854-1860.

Figure 2 Indirect immunofluorescence demonstrating RASA reactivity with SAGA-1 on the surface of human spermatozoa.

The (A) RASA (1:50) and (B) S19 (1:100) mAbs immunolocalized over the entirety of paraformaldehyde-fixed, air-dried spermatozoa. Only faint background fluorescence was observed with the secondary antibodies alone.

Norton, E. J., Diekman, A. B., Westbrook, V. A., Flickinger, C. J., & Herr, J. C. (2001). RASA, a recombinant single-chain variable fragment (scFv) antibody directed against the human sperm surface: implications for novel contraceptives. Human Reproduction, 16(9), 1854-1860.

Figure 3 Agglutination of human spermatozoa with RASA.

RASA or the S19 mAb was mixed with human spermatozoa at a concentration of 20×10⁶ spermatozoa/ml and 1:10 dilution of antibody. After 2 min, sperm motility was observed and recorded under DIC microscopy. RASA alone (A), RASA with anti-E Tag antibody (1:100), (B), and the S19 mAb positive control (C). Spermatozoa were agglutinated in a head-to-head, head-to-tail, and tail-to-tail pattern. Agglutination was not seen with the inactive scFv and anti-E Tag antibody (D), or in samples with anti-E Tag antibody alone.

Norton, E. J., Diekman, A. B., Westbrook, V. A., Flickinger, C. J., & Herr, J. C. (2001). RASA, a recombinant single-chain variable fragment (scFv) antibody directed against the human sperm surface: implications for novel contraceptives. Human Reproduction, 16(9), 1854-1860.