Figure 1

Characterization of gemin antibodies by western blots of total HeLa cell proteins. An SDS extract of HeLa cells was run as a horizontal strip alongside Mr markers on SDS-PAGE and transferred to nitrocellulose membranes by electroblotting. 7% acrylamide was used for (a)-(c) with prestained Sigma markers and 14% for (d) and (e) with prestained SeeBlue markers. The blot was either cut into vertical lanes or, in (c) only, used directly on a 28-lane miniblotter apparatus. In each case, the main antigen band is shown as a broken arrow. (a) gemin3 antiserum stains a single protein of about 102 kD (97 kD gemin4 is shown as a marker). (b) the panel of 6 gemin4 mAbs all stain a band of 97 kD, but 3 of them (GEM4A, C and F) cross-react with several higher Mr proteins. Non-specific bands (*) are due to the secondary antibody system. (c) the 12 mAbs against the gemin5 N-terminal region stain a single band consistent with the 167 kD expected for gemin5 and the 6 mAbs against the C-terminal region stain the same band, together with a ladder of smaller bands that may be degradation products. (d) The 7 gemin6 mAbs all stain a band of 16 kD, but 2 of them cross-react with a 32 kD band. (e) All 7 gemin7 mAbs stain a single band of 15 kD.