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Product details for PARP (Human) antibody
PARP (Human) antibody
Overview
| Antigen: | PARP |
| Antibody Type: | Monoclonal |
| Application: | Western Blotting (WB), Immunocytochemistry (ICC), ELISA |
| Reactivity: | Human, Mouse (Murine), Rat, Mink, Hamster |
| Host: | Mouse |
| Quantity: | |
| Price: | 306,00 € (Plus shipping costs and VAT) |
| Order number: | ABIN130757 |
| Availability: | 7 to 10 days until shipment |
Additional Information: PARP (Human) antibody
| Antigen | PARP |
| Immunogen | Immunogen used was purified calf thymus PARP. EPITOPE: The antibody detects the C-terminal DNA binding domain of PARP. |
| Reactivity | Human, Mouse (Murine), Rat, Mink, Hamster |
| Antibody Type | Monoclonal |
| Isotype | IgG1 »Matching secondary antibodies |
| Description | PARP is a highly conserved nuclear enzyme present in higher eukaryotes. The enzyme is a Zn 2+ -dependent DNA binding protein that recognizes DNA strand breaks and is implicated in DNA repair and in apoptosis responses of cells. As a marker for apoptosis, it has been shown that PARP cleavage mediated by caspase-3 activity occurs at the onset of apoptosis, and inhibition of PARP cleavage attenuates apoptosis in vitro. |
| Clone | C-2-10 |
| Host | Mouse |
| Specificity | The antibody recognizes the 116 kDa PARP and the 85 kDa apoptosis-related cleavage fragment. Recognizes PARP in human, mouse, rat, monkey, bovine, and hamster. |
Application Details: PARP (Human) antibody
| Application | Western Blotting (WB), Immunocytochemistry (ICC), ELISA |
| Application Notes | EXTRACTION OF CELLULAR PARP Buffer Preparation - Stock1. HL60 cell extracts were prepared from exponentially growing cells either uninduced or induced for apoptosis with etoposide. Cells were washed once with PBS, suspended at ~4 X 10 6 cells/ml in sample buffer (62.5 mM Tris / HCl, pH 6.8, 6M urea, 10% glycerol, 2% SDS, 0.00125% bromophenol blue, 5% B-mercaptoethanol), sonicated for 15 seconds and incubated at 65 o C for 15 min. Note: The purpose of the urea in the sample buffer and the sonication step in the above protocol is to effectively dissociate PARP / DNA interactions. Research Use ONLY 2. Run SDS-PAGE (10 or 12% acrylamide gel) using 20 µl (20-40 µg total protein) of control and induced HL60 cell extract. 3. Transfer proteins to nitrocellulose (NC) blotting membrane. 4. Block NC with 5% non-fat dry milk in TBST (50 mM Tris / HCl, pH 7.4, 150 mM NaCl, 0.1% Tween 20) for 1 hr at RT with gentle agitation. 5. Incubate NC with appropriate dilution (see data sheet) of JM-3001 in blocking solution for 2.5 hr at RT. 6. Wash membrane with TBST 3x, 10 minutes per wash. 7. Incubate NC with secondary antibody (1/2000 in Blocking solution) using either anti-mouse or anti-rabbit alkaline phosphatase conjugate, as appropriate, for 1 hour at RT. 8. Wash membrane with TBST 3x, 10 minutes per wash. Rinse briefly with TBS (TBST without Tween). 9. Incubate with BCIP / NBT color development reagent until bands are desired darkness. Rinse with TBS plus 20 mM EDTA to stop color development. Note: Color development times may vary. For chemiluminescent detection, substitute PBS-Tween-20 for TBST and substitute appropriate HRP-labeled antibodies in step 7 and chemiluminescent detection reagents in step 9. These procedures are intended only as a guide. The individual user must determine the optimal concentration of primary antibody and the experimental conditions. No warranty or guarantee of performance using these procedures is made or implied. The antibody can be used for Western blotting (1 µg/ml), ELISA (0.1-0.5 µg/ml), immunohisto- chemistry (frozen and formalin-fixed, paraffin-embedded tissue sections) and immuno- cytochemistry. However, the optimal conditions should be determined individually. For Western blotting, sonication and urea are necessary to break up PARP/DNA interactions. To do this, resuspend cells in 62.5 mM Tris-HCl, pH 6.8, 6 M urea, 10% glycerol, 2% SDS, 0.00125% bromophenol blue and 5% ß-mercaptoethanol, sonicate for 15 seconds on full power and incubated at 65 o C for 15 minutes, and then follow standard Western blotting procedures. Untreated cells should show only full-length PARP, while treated cells will show both cleaved and full-length PARP. |
| Buffer | This buffer will be used in the preparation of the experimental buffers. The final composition of this buffer is: 100mM Tris-HCl (pH 7.4), 10mM MgSO 4 -7H 2 O, 500 mM Sucrose. To make 100 ml of Stock Buffer mix the following: 10.0 ml 1M Tris-HCl (pH 7.4) |
| Research Area | Enzymes, Metabolism |
| Restrictions | For Research Use Only |
Publications: PARP (Human) antibody
| Publications |
Nicholson, Ali, Thornberry et al.: "Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis." In: Nature , Vol. 376, Issue 6535, pp. 37-43, 1995/01 (PubMed) Shah, Shah, Poirier: "Different cleavage pattern for poly(ADP-ribose) polymerase during necrosis and apoptosis in HL-60 cells." In: Biochemical and biophysical research communications , Vol. 229, Issue 3, pp. 838-44, 1997/01 (PubMed) |
External Information: PARP (Human) antibody
| Google Scholar | Find more references for clone C-2-10 on Google Scholar™ |
| EMBL Harvester | Search human, mouse or rat genome for antigen PARP (Bioinformatic Harvester (EMBL)) |
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