The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 98, 29 kDa (predicted molecular weight: 25 kDa).
Involved in the base excision repair (BER) pathway, by catalyzing the poly(ADP-ribosyl)ation of a limited number of acceptor proteins involved in chromatin architecture and in DNA metabolism. This modification follows DNA damages and appears as an obligatory step in a detection/signaling pathway leading to the reparation of DNA strand breaks. Mediates the poly(ADP-ribosyl)ation of APLF and CHFR. Positively regulates the transcription of MTUS1 and negatively regulates the transcription of MTUS2/TIP150. With EEF1A1 and TXK, forms a complex that acts as a T-helper 1 (Th1) cell-specific transcription factor and binds the promoter of IFN-gamma to directly regulate its transcription, and is thus involved importantly in Th1 cytokine production. Required for PARP9 and DTX3L recruitment to DNA damage sites. PARP1-dependent PARP9-DTX3L-mediated ubiquitination promotes the rapid and specific recruitment of 53BP1/TP53BP1, UIMC1/RAP80, and BRCA1 to DNA damage sites.
Phosphorylated by PRKDC and TXK. Poly-ADP-ribosylated by PARP2. Poly-ADP-ribosylation mediates the recruitment of CHD1L to DNA damage sites. S-nitrosylated, leading to inhibit transcription regulation activity.
Nucleus. Nucleus, nucleolus. Localizes at sites of DNA damage.
Western blot - Anti-Cleaved PARP1 antibody [E51] (HRP) (ab194217)
All lanes : Anti-Cleaved PARP1 antibody [E51] (HRP) (ab194217) at 1/1000 dilution
Lane 1 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 20 µg Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate - Treated with 10µM Camptothecin at 20 µg
Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 25 kDa Additional bands at : 29 kDa (possible cleavage fragment),98 kDa (possible immature (unprocessed)).
Exposure time : 8 minutes
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab194217 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.