Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Flow cytometry [1]
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Validation data
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- Product number
- 12-6668-41 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-PARP1 (cleaved Asp214) Monoclonal Antibody (HLNC4), PE, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: This HLNC4 monoclonal antibody reacts with human poly (ADP-ribose) polymerase (PARP1). This ubiquitous 116 kDa nuclear enzyme is involved in DNA repair. During apoptosis, active caspases -3, -6 and -7 cleave PARP1 after Asp214, thereby inactivating PARP1 and generating two apoptotic fragments sized 85 kDa and 25 kDa. The HLNC4 antibody specifically recognizes the 85 kDa PARP1 fragment produced after cleavage and does not recognize the full-length 116 kDa protein.The following peptide was used as the immunogen: NH2-GVDEVAKKKSKKEKDC-COOH. Applications Reported: This HLNC4 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This HLNC4 antibody has been pre-titrated and tested by flow cytometric analysis of staurosporine-stimulated Jurkat cells using the Foxp3/Transcription Factor Staining Buffer Set (cat. 00-5523) and protocol. (Refer to Protocol B: One step protocol for intracellular (nuclear) proteins). This can be used at 5 µL (0.004 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Mouse
- Conjugate
- Yellow dye
- Isotype
- IgG
- Antibody clone number
- HLNC4
- Vial size
- 25 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Jurkat cells were left unstimulated (left) or stimulated for 20 hours with Anti-Human CD95 (APO-1/Fas) Functional Grade Purified (Product # 16-0958-81) coated at 5 µg/mL in a 24-well culture plate (right). The stimulated cells were then harvested and stained sequentially with Fixable Viability Dye eFluor® 780 (Product # 65-0865-14) and the Annexin V Apoptosis Detection Kit eFluor® 450 (Product # 88-8006-72), then fixed and permeabilized with the Foxp3/transcription Factor Staining Buffer Set (Product # 00-5523-00) and stained with Anti-Human PARP1 (Cleaved) PE. Total viable cells (Fixable Viability Dye eFluor® 780 negative) were used for analysis.
- Conjugate
- Yellow dye