1. p100: A Novel Proliferation-Associated Nuclear Protein Specifically Restricted to Cell Cycle Phases S, G2 , and M
by H.J. Heidebrecht, F. Buck, J. Steinmann, R. Sprenger, H.H. Wacker, and R. Parwaresch
Blood
Volume 90(1):
July 1, 1997
©1997 by American Society of Hematology

2. Immunohistochemical staining of a microwave-processed paraffin section of a formalin-fixed human tonsil with Ki-S2.
Immunohistochemical staining of a microwave-processed paraffin section of a formalin-fixed human tonsil with Ki-S2. Cell nuclei in the dark zone of a germinal center, which harbors proliferating cells, are strongly positive. APAAP staining.
H.J. Heidebrecht et al. Blood 1997;90:
©1997 by American Society of Hematology

3. Flow cytometric analysis of unstimulated (day 0) and PHA-stimulated PBMC (days 1 through 3).
Flow cytometric analysis of unstimulated (day 0) and PHA-stimulated PBMC (days 1 through 3). DNA staining was registered on a linear scale (FL1-A) and Ki-S2 staining on a log scale (FL2-H). Less than 1% of the unstimulated PBMC were positive. Twenty-four hours after PHA stimulation, a small population of the G1 cells became positive. On days 2 and 3, all cells in S/G2 and M phases bound Ki-S2.
H.J. Heidebrecht et al. Blood 1997;90:
©1997 by American Society of Hematology

4. Staining of L428 cells with Ki-S2 and a Cy3-labeled goat antimouse polyclonal antibody.
Staining of L428 cells with Ki-S2 and a Cy3-labeled goat antimouse polyclonal antibody. During the different phases of mitosis, the Ki-S2 antigen is strongly associated with the spindle poles and the mitotic spindle, whereas in the S and G2 phases, cells the antigen is diffusely distributed throughout the cell nucleus.
H.J. Heidebrecht et al. Blood 1997;90:
©1997 by American Society of Hematology

5. Western blot of lysed L428 cells using the MoAb Ki-S2 (B) after SDS-PAGE (gradient gel 5% to 10%).
Western blot of lysed L428 cells using the MoAb Ki-S2 (B) after SDS-PAGE (gradient gel 5% to 10%). Ki-S2 detects a protein of about 100 kD. (A) Control experiment with an MoAb of the same isotype (IgG1) specific for topoisomerase-IIα (170 kD). (C) Control without primary antibody. Molecular weight standards are shown on the right (in kilodaltons).
H.J. Heidebrecht et al. Blood 1997;90:
©1997 by American Society of Hematology

6. Immunoprecipitation experiment with Ki-S2 on L428 cells after the cells were labeled with [32P]orthophosphate (A).
Immunoprecipitation experiment with Ki-S2 on L428 cells after the cells were labeled with [32P]orthophosphate (A). In lane B, L428 cells were arrested with colcemid (0.15 μg/mL) overnight before labeling with [32P]. (C) Isotype control experiment. Molecular weight standards are shown on the right (in kilodaltons).
H.J. Heidebrecht et al. Blood 1997;90:
©1997 by American Society of Hematology

7. Determination of the turnover time of the Ki-S2 antigen.
Determination of the turnover time of the Ki-S2 antigen. L428 cells were labeled for 1 hour with [35S]methionine and intensively washed. Immunoprecipitation experiments were performed immediately after the end of labeling (A) and 90 minutes (B), 120 minutes (C), and 180 minutes (D) after the end of labeling. (E) Isotype control experiment after the end of labeling. Molecular weight standards are shown on the right (in kilodaltons).
H.J. Heidebrecht et al. Blood 1997;90:
©1997 by American Society of Hematology

8. A Coomassie blue staining of a PVDF membrane after an immunoprecipitation experiment using cell nuclei of 1 × 1010 HeLa cells with Ki-S2.
A Coomassie blue staining of a PVDF membrane after an immunoprecipitation experiment using cell nuclei of 1 × 1010 HeLa cells with Ki-S2. The 100-kD protein band of this experiment was excised and used for digestion with Lys C and peptide sequencing. Molecular weight standards are shown on the right (in kilodaltons).
H.J. Heidebrecht et al. Blood 1997;90:
©1997 by American Society of Hematology