Figure 1 Western-Blot analysis of HPV-18 E7 protein. WB was carried out with 10A8 Mab using the lysates of U20S, Hela and SiHa cells. Line 1: U20S with plasmid expressing E7 protein, Line 2. Non-transfectedplain U2OS cells. Lines 3 and 4. Lysate of Hela and SiHa cells accordingly. The band in the 3rd lane represents positive control (Hela contains integrated HPV-18). Line 5. Protein size marker.
Figure 2. IP analysis of HPV-18 E7 protein. Line 1. detection of E7 protein by Mab 10A8 in lysate of U20S cells. Line 2. the lower band is precipitated E7, upper bands are light and heavy chain of Mab. Line 3. precipitation without mab 10A8. Line 4. IP exp. with non-transfected U2OS cells. Line 5. IP exp. Non-transfected Hela cells (Hela contains integrated HPV-18). Line 6. Protein size marker.
Figure 3. IF analysis of HPV-18 E7 protein in U2OS cells. IF was carried out with U2OS cells transfected with plasmid expressing HPV-18 E7 protein. All the nuclei were stained by DAPI. Green color represents nucleus-localized HPV-18
: ELISA: 1: 256 000, WB: 1: 1000, IP 1:500.Monoclonal antibody working titer has to be established practically for each particular antigen and assay format