Anti-PARP1 antibody (ab227244)
Overview
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Product name
Anti-PARP1 antibody
See all PARP1 primary antibodies -
Description
Rabbit polyclonal to PARP1 -
Host species
Rabbit -
Tested applications
Suitable for: ChIP, IHC-P, WB, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment within Human PARP1 (internal sequence). The exact sequence is proprietary.
Database link: P09874 -
Positive control
- WB: HEK-293T, NIH/3T3 and PC-12 whole cell extracts; HCT 116, HEK-293T, A431, HeLa and HepG2 whole cell lysate (ab7900). ICC/IF: HeLa cells. ChIP: HeLa and Raji chromatin extracts. IP: HCT 116 whole cell extract. IHC-P: HeLa xenograft tissue.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.00
Preservative: 0.025% Proclin 300
Constituents: PBS, 1% BSA, 20% Glycerol -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
Our Abpromise guarantee covers the use of ab227244 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ChIP | Use at an assay dependent concentration. | |
| IHC-P | 1/100 - 1/1000. | |
| WB | 1/500 - 1/10000. Predicted molecular weight: 113 kDa. | |
| ICC/IF | 1/100 - 1/1000. | |
| IP | 1/100 - 1/500. |
Target
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Function
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. -
Sequence similarities
Contains 1 BRCT domain.
Contains 1 PARP alpha-helical domain.
Contains 1 PARP catalytic domain.
Contains 2 PARP-type zinc fingers. -
Post-translational
modificationsPhosphorylated by PRKDC. Phosphorylated upon DNA damage, probably by ATM or ATR.
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. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 142 Human
- Entrez Gene: 11545 Mouse
- Entrez Gene: 25591 Rat
- Omim: 173870 Human
- SwissProt: P09874 Human
- SwissProt: P11103 Mouse
- SwissProt: P27008 Rat
- Unigene: 177766 Human
see all -
Alternative names
- ADP ribosyltransferase (NAD+; poly (ADP ribose) polymerase) antibody
- ADP ribosyltransferase antibody
- ADP ribosyltransferase diphtheria toxin like 1 antibody
see all
Images
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Western blot - Anti-PARP1 antibody (ab227244)All lanes : Anti-PARP1 antibody (ab227244) at 1/5000 dilution
Lane 1 : Non-transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell extract
Lane 2 : PARP1 shRNA transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell extract
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG
Predicted band size: 113 kDa7.5% SDS-PAGE gel.
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Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody (ab227244)4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for PARP1 (green) using ab227244 at 1/500 dilution in ICC/IF.
Red: Phalloidin, a cytoskeleton marker, at 1/200 dilution.
Blue: Hoechst 33342 staining.
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ChIP - Anti-PARP1 antibody (ab227244)ChIP was performed with HeLa chromatin extract and 5 µg of either normal rabbit IgG or ab227244. The precipitated DNA was detected by PCR with primer set targeting to HSP70.1 promoter.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody (ab227244)Paraffin-embedded HeLa xenograft tissue stained for PARP1 using ab227244 at 1/500 dilution in immunohistochemical analysis.
Antigen Retrieval: EDTA based, pH 8.0, buffer, 15minutes.
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Immunoprecipitation - Anti-PARP1 antibody (ab227244)PARP1 was immunoprecipitated from HCT 116 (human colorectal carcinoma cell line) whole cell extract with 4 µg ab227244. Western blot was performed from the immunoprecipitate using ab227244 at 1/500 dilution. Anti-Rabbit IgG was used as a secondary reagent.
Lane 1: HCT 116 whole cell extract 30 μg.
Lane 2: Control IP in HCT 116 whole cell extract with 4 μg of preimmune Rabbit IgG.
Lane 3: ab227244 IP in HCT 116 whole cell extract.
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Western blot - Anti-PARP1 antibody (ab227244)All lanes : Anti-PARP1 antibody (ab227244) at 1/1000 dilution
Lane 1 : 293T whole cell extracts
Lane 2 : A431 whole cell extracts
Lane 3 : HeLa whole cell extracts
Lane 4 : HepG2 whole cell extracts
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Rabbit IgG antibody (HRP) at 1/10000 dilution
Predicted band size: 113 kDa5% gel.
Running conditions: 80V, 15min; 140V, 40 minutes.
Transfer condition: Semi-dry, 18 V, 60 min (NC membrane).
Blocking condition: 5% non-fat milk in TBST, RT, 60 minutes.
Primary antibody incubation: 4? , overnight.
Washing condition: 5 ml TBST, 4 x 5 minutes.
Exposure: chemiluminescent substrate for the detection of HRP-conjugated antibody
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Western blot - Anti-PARP1 antibody (ab227244)All lanes : Anti-PARP1 antibody (ab227244) at 1/1000 dilution
All lanes : Neuro2A whole cell extracts
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Rabbit IgG antibody (HRP) at 1/10000 dilution
Predicted band size: 113 kDa5% gel.
Running conditions: 80V, 15min; 140V, 40 minutes.
Transfer condition: Semi-dry, 18 V, 60 min (NC membrane).
Blocking condition: 5% non-fat milk in TBST, RT, 60 minutes.
Primary antibody incubation: 4? , overnight.
Washing condition: 5 ml TBST, 4 x 5 minutes.
Exposure: chemiluminescent substrate for the detection of HRP-conjugated antibody
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Western blot - Anti-PARP1 antibody (ab227244)All lanes : Anti-PARP1 antibody (ab227244) at 1/5000 dilution
Lane 1 : HCT116 whole cell lysate (untreated)
Lane 2 : HCT116 whole cell lysate (30 µM cisplatin treatment for 24 hours)
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG
Predicted band size: 113 kDa7.5% SDS-PAGE gel.
The top band is full-length and the bottom band on lane 2 is cleaved PARP1.
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ChIP - Anti-PARP1 antibody (ab227244)Cross-linked ChIP was performed with Raji chromatin extract and 5 μg of either control rabbit IgG or ab227244. The precipitated DNA was detected by PCR with primer set targeting to S100A9 promoter.
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Western blot - Anti-PARP1 antibody (ab227244)All lanes : Anti-PARP1 antibody (ab227244) at 1/500 dilution
Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell extract
Lane 2 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell extract
Secondary
All lanes : HRP-conjugated anti-rabbit IgG
Predicted band size: 113 kDa7.5% SDS-PAGE gel.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
References
ab227244 has been referenced in 3 publications.
- Hughey CC et al. Dysregulated transmethylation leading to hepatocellular carcinoma compromises redox homeostasis and glucose formation. Mol Metab 23:1-13 (2019). PubMed: 30850319
- Dok R et al. Radiosensitization approaches for HPV-positive and HPV-negative head and neck squamous carcinomas. Int J Cancer N/A:N/A (2019). PubMed: 31283004
- Yu Z et al. Bufalin suppresses hepatocarcinogenesis by targeting ß-catenin/TCF signaling via cell cycle-related kinase. Sci Rep 8:3891 (2018). PubMed: 29497076
