Product Pathways - PathScan ELISA
PathScan® Cleaved PARP (Asp214) Sandwich ELISA Kit #7262
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|Kit Includes||Volume||Solution Color|
|Cleaved-PARP (Asp214) Antibody Coated Microwells||96 tests|
|Biotinylated PARP Detection Rabbit mAb||11 ml||Green|
|HRP-linked Streptavidin||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
CST's PathScan® Cleaved PARP (Asp214) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of cleaved PARP (Asp214) protein. A cleaved PARP (Asp214) Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, cleaved PARP (Asp214) protein is captured by the coated antibody. Following extensive washing, a biotinylated PARP Rabbit mAb is added to detect the captured cleaved PARP protein. HRP-linked Streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of cleaved PARP (Asp214) protein.Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
CST's PathScan® Cleaved PARP (Asp214) Sandwich ELISA Kit detects endogenous levels of cleaved PARP (Asp214) protein. As shown in Figure 1, using Cleaved PARP (Asp214) ELISA Kit #7262, a significant induction of cleaved PARP (Asp214) is detected in HeLa cells treated with staurosporine. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
ELISA - Western correlation
Figure 1: Treatment of HeLa cells with staurosporine stimulates cleavage of PARP at Asp214 as detected by PathScan® Cleaved PARP (Asp214) Sandwich ELISA Kit #7262. Absorbance at 450 nm is shown in the top figure, while the corresponding Western blot using Cleaved PARP (Asp214) Antibody #9548 (left panel) or PARP Antibody #9542 (right panel) is shown in the bottom figure. #9542 detects both full-length and cleaved PARP.
Figure 2: The relationship between protein concentration of lysates from untreated and staurosporine (STO) treated HeLa cells and the absorbance at 450 nm is shown. HeLa cells (80% confluent) were treated with staurosporine (1 µM) for 3 hours. Absorbance at 450 nm is shown in the top figure, while the corresponding Western blot using Cleaved PARP Antibody #9548 is shown in the bottom figure.
PARP, a 116 kDa nuclear poly (ADP-ribose) polymerase, appears to be involved in DNA repair in response to environmental stress (1). This protein can be cleaved by many ICE-like caspases in vitro (2,3) and is one of the main cleavage targets of caspase-3 in vivo (4,5). In human PARP, the cleavage occurs between Asp214 and Gly215, which separates the PARP amino-terminal DNA binding domain (24 kDa) from the carboxy-terminal catalytic domain (89 kDa) (2,4). PARP helps cells to maintain their viability; cleavage of PARP facilitates cellular disassembly and serves as a marker of cells undergoing apoptosis (6).
- Satoh, M.S. and Lindahl, T. (1992) Nature 356, 356-358.
- Lazebnik, Y. A. et al. (1994) Nature 371, 346-347.
- Cohen, G.M. (1997) Biochem. J. 326, 1-16.
- Nicholson, D. W. et al. (1995) Nature 376, 37-43.
- Tewari, M. et al. (1995) Cell 81, 801-809.
- Oliver, F.J. et al. (1998) J. Biol. Chem. 273, 33533-33539.
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- 9546 Cleaved PARP (Asp214) (19F4) Mouse mAb (Human Specific)
- 9541 Cleaved PARP (Asp214) Antibody (Human Specific)
- 9548 Cleaved PARP (Asp214) (7C9) Mouse mAb (Mouse Specific)
- 9544 Cleaved PARP (Asp214) Antibody (Mouse Specific)
- 9545 Cleaved PARP (Asp214) Antibody (Rat Specific)
- 9532 PARP (46D11) Rabbit mAb
- 9542 PARP Antibody
- 9929 Cleaved Caspase Antibody Sampler Kit
- 9915 Apoptosis Antibody Sampler Kit
- 9930 Apoptosis Antibody Sampler Kit (Mouse Preferred)
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7004 TMB Substrate
- 7002 STOP Solution
Rabbit Monoclonals are produced under license (granting certain rights, including those under U.S. Patent No. 5,675,063) from Epitomics, Inc.
For Research Use Only. Not For Use In Diagnostic Procedures.