Recombinant Mouse Antibody (M75) is capable of binding to CA9. The adhesion site of CA IX appears to overlap with the PGEEDLP epitope of the monoclonal antibody M75 since the antibody blocks attachment of cells to immobilized CA IX protein.
Figure 1 Strong membrane-stained CA-9 expression (×200).
CA-9 staining was present in 74 of the 101 cancers (73.3%) with no positive staining in any of the control sections.
Rasheed, S., Harris, A. L., Tekkis, P. P., Turley, H., Silver, A., McDonald, P. J., ... & Guenther, T. (2009). Assessment of microvessel density and carbonic anhydrase-9 (CA-9) expression in rectal cancer. Pathology-Research and Practice, 205(1), 1-9.
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 5 The double immunofluorescence staining
The double immunofluorescence images of CA IX (green) and CA II (red) in the human (D) gastric pit and (E ) gastric gland areas show that both proteins occur together in the (D) surface epithelial cells and (E ) parietal cells (arrows). The yellow indicates a colocalization of the proteins.
Pastorekova, S. I. L. V. I. A., Parkkila, S. E. P. P. O., Parkkila, A. K., Opavsky, R. E. N. É., Zelnik, V., Saarnio, J., & Pastorek, J. (1997). Carbonic anhydrase IX, MN/CA IX: analysis of stomach complementary DNA sequence and expression in human and rat alimentary tracts. Gastroenterology, 112(2), 398-408.
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CAT | Product Name | Application | Type |
---|---|---|---|
TAB-707 | Anti-CAIX/CA9 Recombinant Antibody (Girentuximab) | WB, FuncS, IF, Neut, ELISA, FC, IP | IgG1 - kappa |
TAB-H43 | Anti-Human CA9 Recombinant Antibody (Iodine (124I) Girentuximab) | ELISA, IP, IF, Inhib | IgG1 - kappa |
CAT | Product Name | Application | Type |
---|---|---|---|
PFBZ-017 | Mouse Anti-CA9 Recombinant Antibody (clone M75); Fab Fragment | WB, FuncS | Mouse IgG2b |
HPAB-0104-LSX-F(E) | Human Anti-CA9 Recombinant Antibody; Fab Fragment (HPAB-0104-LSX-F(E)) | ELISA | Human Fab |
HPAB-0105-LSX-F(E) | Human Anti-CA9 Recombinant Antibody; Fab Fragment (HPAB-0105-LSX-F(E)) | ELISA | Human Fab |
HPAB-0782-CN-F(E) | Human Anti-CA9 Recombinant Antibody (clone G119); Fab Fragment | ELISA, FC | Human Fab |
HPAB-0784-CN-F(E) | Human Anti-CA9 Recombinant Antibody (clone G37); Fab Fragment | ELISA, FC | Human Fab |
CAT | Product Name | Application | Type |
---|---|---|---|
TAB-1454CL | Anti-Human CA9 Recombinant Antibody (G250) | Cyt | |
TAB-1455CL | Anti-Human CA9 Recombinant Antibody (S239D/I332E) | SPR | |
TAB-1456CL | Anti-Human CA9 Recombinant Antibody (MSC1) | ELISA, FC, Inhib | Fully human antibody |
TAB-1457CL | Anti-Human CA9 Recombinant Antibody (MSC2) | ELISA, FC, Inhib | Fully human antibody |
TAB-1459CL | Anti-Human CA9 Recombinant Antibody (MSC4) | ELISA, FC, Inhib | Fully human antibody |
CAT | Product Name | Application | Type |
---|---|---|---|
MOR-0427 | Hi-Affi™ Rabbit Anti-CA9 Recombinant Antibody (clone DS427AB) | IHC-P, WB | Rabbit IgG |
CAT | Product Name | Application | Type |
---|---|---|---|
HPAB-0104-LSX | Human Anti-CA9 Recombinant Antibody (HPAB-0104-LSX) | ELISA, FC, Inhib | Human IgG |
HPAB-0105-LSX | Human Anti-CA9 Recombinant Antibody (HPAB-0105-LSX) | ELISA, FC, Inhib | Human IgG |
HPAB-0782-CN | Human Anti-CA9 Recombinant Antibody (clone G119) | FC, ELISA, Inhib, Cyt, Bioimaging | Human IgG |
VS3-QX140 | Mouse Anti-CA9 Recombinant Antibody (clone 5B5-B10-A2) | WB, IHC, ICC, FC, ELISA | Mouse IgG1 |
VS4-WK54 | Mouse Anti-CA9 Recombinant Antibody (VS4-WK54) | ELISA, WB, IP | Mouse IgG |
CAT | Product Name | Application | Type |
---|---|---|---|
HPAB-0104-LSX-S(P) | Human Anti-CA9 Recombinant Antibody; scFv Fragment (HPAB-0104-LSX-S(P)) | ELISA | Human scFv |
HPAB-0105-LSX-S(P) | Human Anti-CA9 Recombinant Antibody; scFv Fragment (HPAB-0105-LSX-S(P)) | ELISA | Human scFv |
HPAB-0355-CN-S(P) | Mouse Anti-CA9 Recombinant Antibody; scFv Fragment (HPAB-0355-CN-S(P)) | WB, ELISA, FC | Mouse scFv |
HPAB-0356-CN-S(P) | Mouse Anti-CA9 Recombinant Antibody; scFv Fragment (HPAB-0356-CN-S(P)) | WB, ELISA, FC | Mouse scFv |
HPAB-0357-CN-S(P) | Mouse Anti-CA9 Recombinant Antibody; scFv Fragment (HPAB-0357-CN-S(P)) | WB, ELISA, FC | Mouse scFv |
CAT | Product Name | Application | Type |
---|---|---|---|
AFC-TAB-H43 | Afuco™ Anti-CA9 ADCC Recombinant Antibody (Girentuximab), ADCC Enhanced | ELISA, FC, IP, FuncS, IF, Neut | ADCC enhanced antibody |
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