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Summarized data on the characterization of the monoclonal antibodies (MAbs) raised against carbonic anhydrase XII HEK (CA XII HEK ) using ELISA, Western blot (WB), and flow cytometry (FC) assays.

Journal: International Journal of Molecular Sciences

Article Title: Inhibitory Monoclonal Antibodies and Their Recombinant Derivatives Targeting Surface-Exposed Carbonic Anhydrase XII on Cancer Cells

doi: 10.3390/ijms21249411

Figure Lengend Snippet: Summarized data on the characterization of the monoclonal antibodies (MAbs) raised against carbonic anhydrase XII HEK (CA XII HEK ) using ELISA, Western blot (WB), and flow cytometry (FC) assays.

Article Snippet: MAbs (different mouse IgG subtypes) were purified from hybridoma supernatants, and the scFv-Fc (human IgG1) fusion protein was purified from CHO supernatant using rProtein A Sepharose Fast Flow (GE Healthcare Life Sciences, 17127901, Chicago, IL, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Flow Cytometry

( A ) Reactivity of the MAbs with SDS-denatured recombinant CA XII DE3 and CA XII HEK . Left panel: SDS-PAGE; right panels: immunoblot with MAbs 13G2 and 15H1. M—PageRuler Prestained Protein Ladder, 10 to 180 kDa (Thermo Fisher Scientific, Vilnius, Lithuania, 26616). The lysate of HEK-293 cells was used as a negative control. ( B ) Binding of the MAbs with native CA XII localized on the surface of live A498 cells was investigated by flow cytometry. The MAbs 1B10 and 14D6 reactive with cellular CA XII and the non-reactive MAb 9B9 are shown. Isotype control—the in house produced MAb of the relevant isotype raised against Porcine Parvovirus . Secondary antibody: Alexa Fluor 488-conjugated Goat anti-Mouse IgG (H + L) (Thermo Fisher Scientific, A–11029).

Journal: International Journal of Molecular Sciences

Article Title: Inhibitory Monoclonal Antibodies and Their Recombinant Derivatives Targeting Surface-Exposed Carbonic Anhydrase XII on Cancer Cells

doi: 10.3390/ijms21249411

Figure Lengend Snippet: ( A ) Reactivity of the MAbs with SDS-denatured recombinant CA XII DE3 and CA XII HEK . Left panel: SDS-PAGE; right panels: immunoblot with MAbs 13G2 and 15H1. M—PageRuler Prestained Protein Ladder, 10 to 180 kDa (Thermo Fisher Scientific, Vilnius, Lithuania, 26616). The lysate of HEK-293 cells was used as a negative control. ( B ) Binding of the MAbs with native CA XII localized on the surface of live A498 cells was investigated by flow cytometry. The MAbs 1B10 and 14D6 reactive with cellular CA XII and the non-reactive MAb 9B9 are shown. Isotype control—the in house produced MAb of the relevant isotype raised against Porcine Parvovirus . Secondary antibody: Alexa Fluor 488-conjugated Goat anti-Mouse IgG (H + L) (Thermo Fisher Scientific, A–11029).

Article Snippet: MAbs (different mouse IgG subtypes) were purified from hybridoma supernatants, and the scFv-Fc (human IgG1) fusion protein was purified from CHO supernatant using rProtein A Sepharose Fast Flow (GE Healthcare Life Sciences, 17127901, Chicago, IL, USA).

Techniques: Recombinant, SDS Page, Western Blot, Negative Control, Binding Assay, Flow Cytometry, Produced

Primers for amplification of Ig VH and VL regions. H—heavy chain, Lκ—light kappa chain.

Journal: International Journal of Molecular Sciences

Article Title: Inhibitory Monoclonal Antibodies and Their Recombinant Derivatives Targeting Surface-Exposed Carbonic Anhydrase XII on Cancer Cells

doi: 10.3390/ijms21249411

Figure Lengend Snippet: Primers for amplification of Ig VH and VL regions. H—heavy chain, Lκ—light kappa chain.

Article Snippet: MAbs (different mouse IgG subtypes) were purified from hybridoma supernatants, and the scFv-Fc (human IgG1) fusion protein was purified from CHO supernatant using rProtein A Sepharose Fast Flow (GE Healthcare Life Sciences, 17127901, Chicago, IL, USA).

Techniques: Amplification, Sequencing