Mouse Anti-G4 DNA Recombinant Antibody (clone 1H6) (CAT#: FAMAB-0145CQ)

Figure 1 The 1H6 antibody generates discrete nuclear foci in human cell lines, which is sensitive to treatment with DNase.

(a) Representative images of immunofluorescence analysis using the 1H6 antibody in human cells (HeLa). Bar represents 50 μm. (b) Fluorescent microscopy analysis of HeLa cells stained with 1H6 (red), DAPI DNA dye (Blue) and anti-fibrillarin (green) before and after treatment of cells with DNase. Digestion with RQ1 DNase and micrococcal nuclease show loss of 1H6 foci. Control experiments stained with rabbit anti-fibrillarin do not show loss of signal on digestion with RQ1 DNase and micrococcal nuclease. Bar represents 5 μm.

Henderson, A., Wu, Y., Huang, Y. C., Chavez, E. A., Platt, J., Johnson, F. B., ... & Lansdorp, P. M. (2013). Detection of G-quadruplex DNA in mammalian cells. Nucleic acids research, 42(2), 860-869.

Figure 2 The 1H6 antibody generates discrete nuclear foci in human cell lines, which is sensitive to treatment with DNase.

Normal human tissue sections were used for immunohistochemical staining with 1H6 antibodies (dark brown).

Henderson, A., Wu, Y., Huang, Y. C., Chavez, E. A., Platt, J., Johnson, F. B., ... & Lansdorp, P. M. (2013). Detection of G-quadruplex DNA in mammalian cells. Nucleic acids research, 42(2), 860-869.

Figure 3 Punctate fine granular staining of chromosomes and DNA fibers with 1H6.

(a) Representative images of paraformaldehyde-fixed HeLa cells previously lyzed with Triton X-100. The1H6 antibody (red) appears to be located on nuclear DNA and DNA fibers (blue fibers stained with DAPI) emanating from the nucleus. Image has DAPI channel amplified to identify DNA fibers more clearly. Bar represents 50 µm. (b) The1H6 antibody shows many discrete foci on metaphase chromosomes of normal mouse emrbryonic stem cells. (c) The 1H6 antibody shows many discrete foci on chromosomes of human metaphase preparations. Arrows point to bright staining near the centromere of chromosome 9. Representative images of immunofluorescence analysis of metaphase preparations using 1H6 antibodies. Bar represents 10 μm.

Henderson, A., Wu, Y., Huang, Y. C., Chavez, E. A., Platt, J., Johnson, F. B., ... & Lansdorp, P. M. (2013). Detection of G-quadruplex DNA in mammalian cells. Nucleic acids research, 42(2), 860-869.

Figure 4 The 1H6 foci decrease in the presence of G4 DNA and increase in the presence of TMP and TMS.

(a) Synthetic tetramolecular G4 DNA (Oxy-2 tetraplex) decreases 1H6 nuclear foci as detected by flow cytometry. Human HeLa cells were prepared and stained with anti-G-quadruplex antibody 1H6 with and without the presence of synthetic tetramolecular G4 DNA. Mean fluorescence was measured by flow cytometry and plotted above. Incubating the 1H6 antibody in the presence of tetramolecular DNA decreases the total mean fluorescence (n> 25 000). (b) Flow cytometry quantitative analysis of HeLa cells show increased mean fluorescence on treatment with TMPyP4. HeLa cells treated with increasing concentrations of TMPyP4 and subsequently stained with the 1H6 antibody. Increased TMPyP4 treatment resulted in increased mean fluorescence detected for 1H6 but not controls. TMPyP4 concentrations (TMP) in µM are listed below (n> 25 000). (c) Flow cytometry analysis of HeLa cells treated with TMPyP4 or no drug control. Log scale differences observed on treatment in cells stained with 1H6 as compared with controls (n> 20 000).

Henderson, A., Wu, Y., Huang, Y. C., Chavez, E. A., Platt, J., Johnson, F. B., ... & Lansdorp, P. M. (2013). Detection of G-quadruplex DNA in mammalian cells. Nucleic acids research, 42(2), 860-869.

Figure 5 Cells defective in FANC J show increased 1H6 fluorescence.

(a) TMS induces elevated numbers of 1H6 foci in FANCJ DT40 chicken cells. FANCJ DT40 cells or cells expressing human FANCJ-WT were either exposed to 5 μM TMS for 2 h or left unexposed, fixed and stained with 1H6 (red) and DAPI (blue). (b) Quantification of the change in 1H6 foci with and without TMS treatment. The graph illustrates the mean ± s.e.m of the G4 foci in each cell. More than 100 cells were scored for each sample.

Henderson, A., Wu, Y., Huang, Y. C., Chavez, E. A., Platt, J., Johnson, F. B., ... & Lansdorp, P. M. (2013). Detection of G-quadruplex DNA in mammalian cells. Nucleic acids research, 42(2), 860-869.