Overview

  • Product name

  • 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

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

  • 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
    modifications

    Phosphorylated 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
  • Database links

  • Alternative names

    • ADP ribosyltransferase (NAD+; poly (ADP ribose) polymerase) antibody
    • ADP ribosyltransferase antibody
    • ADP ribosyltransferase diphtheria toxin like 1 antibody
    • ADP ribosyltransferase NAD(+) antibody
    • ADPRT 1 antibody
    • ADPRT antibody
    • ADPRT1 antibody
    • ARTD1 antibody
    • msPARP antibody
    • NAD(+) ADP ribosyltransferase 1 antibody
    • NAD(+) ADP-ribosyltransferase 1 antibody
    • pADPRT 1 antibody
    • pADPRT-1 antibody
    • pADPRT1 antibody
    • PARP 1 antibody
    • PARP antibody
    • PARP-1 antibody
    • PARP1 antibody
    • PARP1_HUMAN antibody
    • Poly (ADP ribose) polymerase 1 antibody
    • poly (ADP ribose) polymerase family, member 1 antibody
    • Poly (ADP-ribose) polymerase 1 antibody
    • Poly [ADP-ribose] polymerase 1 antibody
    • Poly(ADP ribose) polymerase antibody
    • poly(ADP ribose) synthetase antibody
    • poly(ADP ribosyl)transferase antibody
    • Poly(ADP-ribosyl)transferase antibody
    • Poly[ADP ribose] synthetase 1 antibody
    • Poly[ADP-ribose] synthase 1 antibody
    • PPOL antibody
    • sPARP 1 antibody
    • sPARP1 antibody
    see all

Images

  • 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 kDa



    7.5% SDS-PAGE gel.

  • 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.

  • 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.

  • 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. 

  • 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.

  • 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 kDa



    5% 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

     

  • 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 kDa



    5% 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

     

  • 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 kDa



    7.5% SDS-PAGE gel.

    The top band is full-length and the bottom band on lane 2 is cleaved PARP1. 

  • 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.

  • 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 kDa



    7.5% SDS-PAGE gel.

References

This product has been referenced in:

  • Hughey CC  et al. Dysregulated transmethylation leading to hepatocellular carcinoma compromises redox homeostasis and glucose formation. Mol Metab 23:1-13 (2019). Read more (PubMed: 30850319) »
  • Yu Z  et al. Bufalin suppresses hepatocarcinogenesis by targeting ß-catenin/TCF signaling via cell cycle-related kinase. Sci Rep 8:3891 (2018). Read more (PubMed: 29497076) »
See all 2 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (B-cell lymphocyte whole cell lysate)
Total protein in input
300 µg
Immuno-precipitation step
Protein A
Specification
B-cell lymphocyte whole cell lysate

Abcam user community

Verified customer

Submitted Mar 13 2019

Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse RPE, retina)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris Gel)
Loading amount
30 µg
Specification
mouse RPE, retina
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Mar 04 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (multiple myeoma cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
20 nm Bortezomib for 16 hours
Specification
multiple myeoma cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Aug 17 2018

Application
Western blot
Sample
Human Cell lysate - other (U87MG cells)
Gel Running Conditions
Non-reduced Denaturing
Loading amount
50 µg
Treatment
10Gray Irradiation
Specification
U87MG cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Mr. Matthias Dedobbeleer

Verified customer

Submitted Jul 18 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (fibroblast)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
30 µg
Specification
fibroblast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 26 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (fibroblast)
Permeabilization
Yes - 0.3% Triton X-100 in blocking buffer
Specification
fibroblast
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 15 2018

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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