Mouse Anti-CA9 Recombinant Antibody (clone M75) (CAT#: PABZ-017)

Figure 2 PAGE and Western blot analysis of MN/CA IX protein.

Lanes 1–3 = gel stained with Coomassie Blue; lanes 4–8 = blots developed with mAb M75 and immunoperoxidase. Lane 1 = Mr reference proteins; lane 2 = purified MN/CA IX ∆TM, IC, 1.25 µg; lane 3 = purified MN/CA IX+, 2 µg; lane 4 = purified MN/CA IX ∆TM, IC, 0.125 µg; lane 5 = purified MN/CA IX+, 0.2 µg; lane 6 = extract from HT29 cells, 50 µl; lane 7 = extract from CGL3 cells, 50 µl; lane 8 = extract from HeLa cells, 50 µl

Zavada, J., Zavadova, Z., Pastorek, J., Biesova, Z., Jezek, J., & Velek, J. (2000). Human tumour-associated cell adhesion protein MN/CA IX: identification of M75 epitope and of the region mediating cell adhesion. British journal of cancer, 82(11), 1808.

Figure 3 Competition ELISA with proteins or oligopeptides related to MN/CA IX protein.

Competing proteins/peptides: (A–C) Recombinant proteins. (A) MN/CA IX+; (B) MN/CA IX∆; (C) GST-MN; (D–G) overlapping proteins covering the PG region: (D) GGSSGEDDPLGEEDLPSEEDSPC (aa 51–72); (E) GEEDLPSEEDSPREEDPPGEEDLPGEC (aa 61–85); (F) EDPPGEEDLPGEEDLPGEEDLPEVC (aa 75–98); (G) EVKPKSEEEGSLKLE (aa 97–111); (H–M) dodecapeptides: (H) GEEDLPGEEDLP; (I) EEDLPGEEDLPG; (J) EDLPGEEDLPGE; (K) DLPGEEDLPGEE; (L) LPGEEDLPGEED; (M) PGEEDLPGEEDL; (N, O) oligopeptides 7 + 2 aa: (N) APGEEDLPA; (O) AGEEDLPGA. Other oligopetides tested which proved negative in competition at 100 µg ml–1 were oligopeptides 7 + 2 aa: AEEDLPGEA, AEDLPGEEA, ADLPGEEDA, ALPGEEDLA and all of the 6 + 2 aa permutations (AGEEDLPA, AEEDLPGA, AEDLPGEA, ADLPGEEA, ALPGEEDA and APGEEDLA). In oligopeptides D, E, F and G the C-terminal amino acid was present as acid, in all others as amide

Zavada, J., Zavadova, Z., Pastorek, J., Biesova, Z., Jezek, J., & Velek, J. (2000). Human tumour-associated cell adhesion protein MN/CA IX: identification of M75 epitope and of the region mediating cell adhesion. British journal of cancer, 82(11), 1808.

Figure 4 Blocking of cell attachment by M75

Adhesion of cells to the dots of MN/CA IX protein immobilized in bacteriological Petri dishes. This plasticware (not treated for use with tissue cultures) allows no adhesion of CGL1 cells and a limited adhesion of HeLa. (A) CGL1 cells do attach and spread to the dot of adsorbed MN/CA IX. (B) Treatment of the dot with MAb M75 abrogates its binding capacity. (C) Treatment with MAb M16 (irrelevant for MN/CA IX) has no effect on cell attachment. (D) HeLa cells attach to complete MN/CA IX+. (E) HeLa cells do not attach to truncated MN/CA IX∆

Zavada, J., Zavadova, Z., Pastorek, J., Biesova, Z., Jezek, J., & Velek, J. (2000). Human tumour-associated cell adhesion protein MN/CA IX: identification of M75 epitope and of the region mediating cell adhesion. British journal of cancer, 82(11), 1808.

Figure 1 Anti-CA9 Antibody in WB.

Western blot analysis of PABZ-017 was performed with Human CA9 Protein (His Tag).

Figure 2 Anti-CA9 Antibody in ELISA.

ELISA analysis of PABZ-017 was performed by coating with Human CA9 Protein (His Tag).

Figure 3 Anti-CA9 Antibody in Dot Blot.

Dot Blot analysis of PABZ-017 was performed by coating with Human CA9 Protein (His Tag).