The relationship between protein concentration of lysates from untreated and staurosporine (STO) treated HeLa cells and the absorbance at 450 nm using PathScan® Cleaved PARP (Asp214) Sandwich ELISA Antibody Pair #7858 is shown. HeLa cells (80% confluent) were treated with staurosporine (1 µM) for 3 hours at 37ºC, and then lysed.
Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
CST's PathScan® Cleaved PARP (Asp214) Sandwich ELISA Antibody Pair is offered as an economical alternative to our PathScan® Cleaved PARP (Asp214) Sandwich ELISA Kit #7262. Capture and Detection antibodies (100X stocks) and HRP-conjugated Streptavidin (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The cleaved PARP (Asp214) Rabbit mAb is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by a Biotinylated PARP Detection Rabbit mAb and HRP conjugated Streptavidin. HRP substrate, TMB, is added for color development. 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.
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
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).
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