Carbonic anhydrase antibody
A carbonic anhydrase and antibody technology, applied in the direction of antibodies, antibody medical components, anti-tumor drugs, etc., can solve problems such as adverse side effects
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Embodiment 1
[0096] Example 1 - CA-XII Expression Levels in Different Types of Human Cancers
[0097] CA-XII is highly expressed in different types of human cancers. Overexpression of CA XII is most pronounced in breast cancer as demonstrated by data documented by Bernett et al. (2008) (available from http: / / ist.genesapiens.org).
Embodiment 2-A549
[0098] Immunofluorescence staining of embodiment 2-A549 cells
[0099] A549 cells were fixed with 4% (v / v) paraformaldehyde (in PBS) for 10 minutes at room temperature, washed with PBS and 0.5% (v / v) Triton-X-100 (in PBS), and Incubate with EXO 6A10 antibody for 60 minutes. After washing with PBS and PBS / 0.5% (v / v) Triton-X-100, cells were incubated with a fluorochrome-labeled secondary antibody (mouse anti-rat IgG / Cy3), and then analyzed by a Leica laser scanning microscope. Bound antibodies were visualized. Nuclei were counterstained with DAPI. Therefore, the staining of CA-XII is clearly associated with the cell membrane of A549 cells ( image 3 ,arrow).
Embodiment 3
[0100] Example 3 - Investigated human cancer cell lines express CA-XII
[0101] refer to Figure 4A , most of the constant human cancer cell lines investigated express CA-XII on the cell surface as shown by flow cytometry. A constant human cancer cell line was incubated on ice with EXO6A10 (hybridoma supernatant diluted 1:5 in 2% (v / v) FCS (e.g., fetal calf serum) in PBS) for 15 minutes, Washed three times in PBS / 2% (v / v) FCS, followed by staining with a specific secondary antibody (goat anti-rat IgG / Cy5) for an additional 15 minutes on ice. Binding of 6A10 was then measured by flow cytometry using a Becton Dickinson FACS Calibur device and FlowJo software (Treestar Corporation). An antibody of the same isotype (glutathione-S-transferase) with an irrelevant specificity was used as a control (gray = isotype control; black line = EXO 6A10). No binding to peripheral blood mononuclear cells (PBMCs) and no binding to two melanoma cell lines (not shown) was detectable. Figure 4...
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