ABCC2 p.Leu23Ser
| Predicted by SNAP2: | A: D (71%), C: D (66%), D: D (85%), E: D (85%), F: N (72%), G: D (80%), H: D (85%), I: N (57%), K: D (91%), M: D (66%), N: D (80%), P: D (85%), Q: D (80%), R: D (91%), S: D (80%), T: D (80%), V: N (66%), W: D (85%), Y: D (75%), |
| Predicted by PROVEAN: | A: D, C: D, D: D, E: D, F: N, G: D, H: D, I: N, K: D, M: N, N: D, P: D, Q: D, R: D, S: D, T: D, V: N, W: D, Y: N, |
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[hide] A new monoclonal antibody, P2A8(6), that specifica... Hybrid Hybridomics. 2001;20(5-6):333-41. Connolly L, Moran E, Larkin A, Scheffer G, Scheper R, Sarkadi B, Kool M, Clynes M
A new monoclonal antibody, P2A8(6), that specifically recognizes a novel epitope on the multidrug resistance-associated protein 1 (MRP1), but not on MRP2 nor MRP3.
Hybrid Hybridomics. 2001;20(5-6):333-41., [PMID:11839251]
Abstract [show]
Multidrug resistance (MDR) is a major problem in the chemotherapeutic treatment of cancer. Overexpression of the multidrug resistance-associated protein 1 (MRP1), is associated with MDR in certain tumors. A number of MRP1-specific MAbs, which facilitate both clinical and experimental investigations of this protein, are available. To add to this panel of existing antibodies, we have now generated an additional MRP1-specific monoclonal antibody (MAb), P2A8(6), which detects a unique heat stable epitope on the MRP1 molecule. Female Wistar rats were immunized via footpad injections with a combination of two short synthetic peptides corresponding to amino acids 235-246 (peptide A) and 246-260 (peptide B) of the MRP1 protein. Immune reactive B cells were then isolated from the popliteal lymph nodes for fusion with SP2/O-Ag14 myeloma cells. Resultant hybridoma supernatants were screened for MRP1-specific antibody production. Antibody P2A8(6) was characterized by Western blotting and immunocytochemistry on paired multidrug resistant (MRP1 overexpressing) and sensitive parental cell lines. The antibody detects a protein of 190 kDa in MRP1-expressing cell lines but not in MRP2- or MRP3-transfected cell lines. P2A8(6) stains drug-selected and MRP1-transfected cell lines homogeneously by immunocytochemistry and recognizes MRP1 by immunohistochemistry on formalin-fixed paraffin wax-embedded tissue sections. Peptide inhibition studies confirm that P2A8(6) reacts with peptide B (amino acids 246-260), therefore recognizing a different epitope from that of all currently available MRP1 MAbs. This new MAb, chosen for its specificity to the MRP1 protein, may be a useful addition to the currently available range of MRP1-specific MAbs.
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No. Sentence Comment
43 Control cell lines The multidrug resistant nonsmall cell lung carcinoma cell line, COR-L23R (made resistant by exposure to increasing concentrations of adriamycin(26)) and its drug-sensitive counterpart, COR-L23S, were used as control cells for the presence/absence of MRP1.
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ABCC2 p.Leu23Ser 11839251:43:208
status: NEW44 Overexpression of MRP1 in COR-L23R cells compared with the parental COR-L23S had been confirmed in previous studies using other commercially available antibodies raised against MRP1 (MRPr1, MRPm5, MRPm6,(15) QCRL-1, and QCRL-3(16)) (Alexis Biochemicals, San Diego, CA).
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ABCC2 p.Leu23Ser 11839251:44:72
status: NEW77 P2A8(6) also detects a very faint 190-kDa band on preparationsfrom drug-sensitivecells (COR-L23S) and non- transfectedcells (2008 parental)indicatingan intrinsic basal expression of MRP1.
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ABCC2 p.Leu23Ser 11839251:77:92
status: NEW110 A very low basal level of MRP1 is detected in the sensitive COR-L23S cells.
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ABCC2 p.Leu23Ser 11839251:110:64
status: NEW115 MRP2 is undetectable in the COR-L23R, COR-L23S, HL60ADR, HL60S, DLKP, RPMI 2650, and RPMI 2650 taxol-selected cells.
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ABCC2 p.Leu23Ser 11839251:115:42
status: NEW126 Neither MAb detected MRP1 expression in the COR-L23S, HL60S, DLKP, DLKPA, or RPMI 2650 taxol-selected cell lines.
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ABCC2 p.Leu23Ser 11839251:126:48
status: NEW134 SUMMARY OF MRP1, MRP2, AND MRP3 PROTEIN EXPRESSION LEVELS AS DETERMINED BY WESTERN BLOT ANALYSIS Protein expression levels (as determined by Western blot analysis) MRPr1 M2III-6 M3II-21 Cell line P2A8(6) (MRP1) (MRP2) (MRP3) COR-L23R 1 1 - - COR-L23S 1/2 1/2 - - HL60ADR 1 1 - - HL60S - - - - DLKP - - - - DLKPA - - - - A549 1 1 1 - RPMI 2650 - 1/2 - - Parental RPMI 2650 - - - - Taxol-selected RPMI 2650 1 1 1 - Melphalan-selected Key: -: Undetectable levels; 1: Over-expression; and 1/2: Very low basal levels.
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ABCC2 p.Leu23Ser 11839251:134:246
status: NEW141 MAb P2A8(6) did not detect overexpressionof MRP1 in the sensitiveCOR-L23S, 2008 parental,2008 MRP2, or 2008 MRP3 cell lines but like MRPr1, P2A8(6) did detect a very low basal level of staining in these cell lines.
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ABCC2 p.Leu23Ser 11839251:141:69
status: NEW