WB
Western blot analysis of Cleaved PARP on different lysates with Rabbit anti-Cleaved PARP antibody at 1/2,000 dilution. Lane 1: HeLa whole cell lysate, Lane 2: HeLa treated with 1μM staurosporine for 3 hours whole cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 1 minute 9 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of HeLa cells treated with 1μM staurosporine for 180 minutes labeling Cleaved PARP with Rabbit anti-Cleaved PARP antibody at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cleaved PARP antibody at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.IP
Cleaved PARP was immunoprecipitated from 0.2 mg Daudi cell lysate with Rabbit anti-Cleaved PARP antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Cleaved PARP antibody at 1/500 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Daudi cell lysate (input), Lane 2: Rabbit anti-Cleaved PARP antibody IP in Daudi cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Cleaved PARP antibody in Daudi cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 12 seconds.| Product Name | Cleaved PARP Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Poly(ADP-ribose) polymerase-1 (PARP-1), also designated PARP, is a nuclear DNA-binding zinc finger protein that influences DNA repair, DNA replication, modulation of chromatin structure, and apoptosis. In response to genotoxic stress, PARP-1 catalyzes the transfer of ADP-ribose units from NAD(+) to a number of acceptor molecules including chromatin. PARP-1 recognizes DNA strand interruptions and can complex with RNA and negatively regulate transcription. Actinomycin D- and etoposide-dependent induction of caspases mediates cleavage of PARP-1 into a p89 fragment that traverses into the cytoplasm. Apoptosis-inducing factor (AIF) translocation from the mitochondria to the nucleus is PARP-1-dependent and is necessary for PARP-1-dependent cell death. PARP-1 deficiencies lead to chromosomal instability due to higher frequencies of chromosome fusions and aneuploidy, suggesting that poly(ADP-ribosyl)ation contributes to the efficient maintenance of genome integrity. This antibody recognizes the apoptosis-specific 89 kDa catalytic domain fragment, but it does not recognize the full-length PARP-1 or the 24 kDa DNA binding domain fragment. |
| Immunogen | Synthetic peptide within Human PARP1 aa 200-249 / 1,014. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | ICC/IFIPWB |
| WB:1:500 ICC/IF:1:50-1:250 IP:1-2μg/sample |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | ADP-ribosyltransferase diphtheria toxin-like 1 antibody ADPRT 1 antibody ADPRT antibody ADPRT1 antibody APOPAIN antibody ARTD1 antibody NAD(+) ADP-ribosyltransferase 1 antibody PARP antibody PARP-1 antibody PARP1 antibody PARP1_HUMAN antibody Poly [ADP-ribose] polymerase 1 antibody Poly ADP ribose polymerase 1 antibody Poly[ADP-ribose] synthase 1 antibody PPOL antibody SCA1 antibody |
|---|---|
| Molecular Weight(MW) | 89kDa |
| Cellular Localization | Nucleus. |
| SwissProt ID | P09874 |
|---|
WB
Western blot analysis of Cleaved PARP on different lysates with Rabbit anti-Cleaved PARP antibody at 1/2,000 dilution. Lane 1: HeLa whole cell lysate, Lane 2: HeLa treated with 1μM staurosporine for 3 hours whole cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 1 minute 9 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of HeLa cells treated with 1μM staurosporine for 180 minutes labeling Cleaved PARP with Rabbit anti-Cleaved PARP antibody at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cleaved PARP antibody at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
IP
Cleaved PARP was immunoprecipitated from 0.2 mg Daudi cell lysate with Rabbit anti-Cleaved PARP antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Cleaved PARP antibody at 1/500 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Daudi cell lysate (input), Lane 2: Rabbit anti-Cleaved PARP antibody IP in Daudi cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Cleaved PARP antibody in Daudi cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 12 seconds.| Application Notes | WB:1:500 ICC/IF:1:50-1:250 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
